Non-hormonal steroid modulators of NF-κB for treatment of disease

ABSTRACT

The present invention relates to compounds and methods which may be useful as treatments of neuromuscular diseases such as muscular dystrophy, and as inhibitors of NF-κB for the treatment or prevention of muscular wasting disease, including muscular dystrophy.

This application claims the benefit of priority of U.S. provisionalapplication No. 61/056,715, filed May 28, 2008, the disclosure of whichis hereby incorporated by reference as if written herein in itsentirety.

This invention was made with support from the U.S. Government underContract No. DOD 05118004 and NIH No. 1U54HD053177-01A1. The U.S.Government has certain rights to this invention.

Disclosed herein are new non-hormonal steroid compounds and compositionsand their application as pharmaceuticals for the treatment of disease.Methods of modulation of NF-κB activity in a human or animal subject arealso provided for the treatment of diseases mediated by NF-κB.

Muscular wasting diseases, such as muscular dystrophies, are a group ofdegenerative diseases that culminate in progressive skeletal musclewasting leading to muscle weakness, a high incidence of bone fracture,wheelchair dependence, and in some cases death. Of the musculardystrophies, Duchenne muscular dystrophy is the most severe and mostwidely recognized. Another muscular wasting disease which shows similarsymptoms, although less severe than Duchenne muscular dystrophy, isBecker muscular dystrophy. Even though the defective dystrophin genecausing both Duchenne muscular dystrophy and Becker muscular dystrophyhas been known for over 20 years, a cure is still lacking.

Several catabolic factors that act to destroy host tissues during thecachectic process have been identified. It appears that theoversecretion of inflammatory cytokines, specifically tumor necrosisfactor-alpha (TNF-α), is one of the most likely causes of cachexia.Specifically, TNF-α can mimic most of the abnormalities that occur incachexia such as weight loss, anorexia, increased thermogenesis, changesin lipid metabolism, insulin resistance, and muscle wasting.

Muscle atrophy can also be induced by the loss of innervation or damageto innervation of the muscle tissue. Diseases such as chronic neuropathyand motor neuron disease can cause damage to innervation. Physicalinjury to the nerve can also lead to damage to the innervation of themuscle tissue. Alternatively, muscle atrophy can be the result ofenvironmental conditions such as during spaceflight or as a result ofaging or extended bed rest. Under these environmental conditions, themuscles do not bear the usual weight load, resulting in muscle atrophyfrom disuse. Specifically, during muscle disuse, intracellular processesare activated to induce proteolysis, mainly through the ATP dependentubiquitin proteasome pathway, which regulates the NF-κB pathway.

NF-κB is known to mediate extracellular signals responsible for theinduction of genes involved in pro-inflammatory responses. NF-κB issequestered in the cytoplasm of most non-stimulated cells through anon-covalent interaction with one of several proteins known asinhibitors of kappa-beta (IκBs) (May & Ghosh, (1997) Semin. Cancer.Biol. 8, 63-73; May & Ghosh, (1998) Immunol. Today 19, 80-88; Ghosh etal., (1998) Annu. Rev. Immunol. 16, 225-260). Cellular stimuliassociated with pro-inflammatory responses such as TNF-α activatekinases, which in turn activate NF-κB by phosphorylating IκBs. Thekinases that phosphorylate IκBs are called IκB kinases (IKKs).

Phosphorylation targets IκBs for subsequent ubiquitination anddegradation. This degradation of IκBs reveals the nuclear localizationsignal on NF-κB, allowing nuclear accumulation of activation, whichleads to binding of DNA and control of specific gene expression.Phosphorylation of IκB is therefore an important step in the regulationof NF-κB downstream of many stimuli, although other mechanisms can leadto the activation of functional NF-κB.

The identification and characterization of kinases that phosphorylateIκBs has led to a better understanding of signaling pathways involvingNF-κB activation. Several different subtypes of IKK have been identifiedthus far. IKKα was initially identified as an IκB kinase induced byTNF-α stimulation in HeLa cells (DiDonato et al., (1997) Nature 388,548-554). Another IκB kinase homologous to IKKα was identified, termedIKKβ, and determined to be the major IκB kinase induced following TNFαstimulation (Takeda et al., (1999) Science 284, 313-316; U.S. Pat. No.6,030,834, issued to Pots et al. (2000); U.S. Pat. No. 5,939,302, issuedto Woronicz et al. (1999)). IKKα and IKKβ have an overall homology of52% and a 65% homology in the kinase domain (Zandi et al., (1997) Cell91, 243-252).

IκB protein kinases (IKKs) phosphorylate IκBs at specific serineresidues. Specifically, they phosphorylate serines 32 and 36 of IκBζ(Traenckner et al., (1995) EMBO J. 14, 2876-2883; DiDonato et al.,(1996) Mol. Cell. Biol. 16, 1295-1304). Phosphorylation of both sites isrequired to efficiently target IκBα for degradation. Furthermore,activation of IκKα and IκKβ is usually in response to NF-κB activatingagents including phorbol 12-myristate 13-acetate (PMA),lipopolysaccharide (LPS), interleukin-1 (IL-1), TNF-α, reactive oxygenspecies, and DNA damaging agents. Mutant IKKα and IKKβ, which arecatalytically inactive, can be used to block NF-κB stimulation. IκBkinases are therefore essential in the regulation of NF-κB activationprocesses downstream of inflammatory stimuli. In other pathways, IκBkinases may not be important.

IKKα and IKKβ have distinct structural motifs including an aminoterminal serine-threonine kinase domain separated from a carboxylproximal helix-loop-helix domain by a leucine zipper domain. Thesestructural characteristics are unlike other kinases, and thenon-catalytic domains are thought to be involved in protein-proteininteractions. As such, proteins which bind to IKKs should be capable ofregulating the activity of NF-κB and potentially regulating downstreamevents such as induction of NF-κB. For instance, NEMO (NF-κB EssentialModulator) is a protein which has been identified to bind to IKKs andfacilitate kinase activity (Yamaoke et al., (1998) Cell 93, 1231-1240;Rothwarf et al., (1998) Nature 395, 287-300).

In vivo studies have shown that chronic NF-κB activation is associatedwith muscular wasting diseases such as Duchenne muscular dystrophy, andis further illustrated in US 2007/0225315 (Mar. 15, 2007). Specifically,muscle wasting was largely prevented in subjects that were heterozygousfor the p65/RelA NF-κB subunit. An injection of an NF-κB activationinhibitor peptide was found to inhibit the dystrophic phenotype inaffected mice subjects. Without being bound by a particular theory, itappears that chronic activation of NF-κB is required for the musclewasting symptoms of Duchenne muscular dystrophy. As such, a drug-basedtherapy targeting NF-κB can be an effective strategy to treat Duchennemuscular dystrophy, as well as other forms of muscular wasting diseases.

In general, muscular wasting diseases may be treated in accordance withthe present disclosure with a direct or indirect modulator of NF-κB.Indirect modulators of NF-κB include, for example, inhibitors of IκBkinases (IKKs) such as IKKα inhibitors and IKKβ inhibitors, andinhibitors functioning directly upstream from IKKs in the signalingpathway such as inhibitors of phosphoinositide dependent kinase (PDK)and inhibitors of Akt (also referred to as PKB).

As noted above, one suitable approach for modulating the NF-κB pathwayis by binding to one of the IκB protein kinases (IKKs). By binding theIKKs, phosphorylation of IκBs is blocked and NF-κB cannot be activated.In one embodiment, direct inhibiting compounds of IKK catalytic activitycan be administered for the purpose of blocking the NF-κB pathway andinhibiting a muscular wasting disease. Specifically, inhibitors of IKKαor their enantiomers, analogs, prodrugs, active metabolites, salts,and/or hydrates thereof can be administered to the subject for thepurpose of inhibiting a muscular wasting disease.

Novel compounds and pharmaceutical compositions, certain of which havebeen found to modulate NF-κB have been discovered, together with methodsof synthesizing and using the compounds including methods for thetreatment of NF-κB-mediated diseases in a patient by administering thecompounds.

In certain embodiments, disclosed herein are compounds having structuralFormula I:

or a salt thereof, wherein:

said dashed line indicates an optional double bond;

R₁, R₂, R₃, and R₄ are each independently selected from the groupconsisting of hydrogen, unsubstituted lower alkyl, lower haloalkyl, andhalogen;

R₅ is selected from the group consisting of hydrogen, lower alkyl, aryl,cycloalkyl, heterocycloalkyl, and heteroaryl, said lower alkyl, aryl,cycloalkyl, heterocycloalkyl, and heteroaryl being optionallysubstituted with one or more substituents selected from the groupconsisting of acyl, alkenyl, alkoxy, alkyl, alkynyl, amido, amino, aryl,aryloxy, cycloalkyl, haloalkoxy, haloalkyl, heteroalkyl, heteroaryl,hydroxy, perhaloalkoxy, and thiol;

R₆ is selected from the group consisting of hydrogen, hydroxyl, andlower alkyl, said lower alkyl being optionally substituted with one ormore substituents selected from the group consisting of alkenyl, alkoxy,alkyl, alkynyl, aryl, aryloxy, haloalkoxy, haloalkyl, heteroalkyl,hydroxy, and thiol;

R₇ and R₈ are independently selected from the groups consisting ofhydrogen, unsubstituted C₁₋₃ alkyl, or R₇ and R₈ can be taken togetherto form oxo or C₃₋₆ saturated cycloalkyl; and

R₉ is selected from the group consisting of hydrogen, acyl, and alkyl,said acyl and alkyl being optionally substituted with one or moresubstituents selected from the group consisting of acyl, alkenyl,alkoxy, alkyl, alkylamino, alkylthio, alkynyl, amido, amino, aryl,aryloxy, aroyl, carbamate, carboxyl, cyano, cycloalkyl, halogen,haloalkoxy, haloalkyl, heteroalkyl, heterocycloalkyl, heteroaryl,hydrazinyl, hydroxy, mercaptyl, nitro, oxo, perhaloalkoxy, sulfonate,alkylsulfonyl, N-sulfonamido, S-sulfonamido, and thiol.

In further embodiments, R₇ and R₈ are independently selected from thegroups consisting of hydrogen, unsubstituted C₂-C₃ alkyl, or R₇ and R₈can be taken together to form C₃₋₆ saturated cycloalkyl; R₉ is selectedfrom the group consisting of hydrogen, acyl, and alkyl, said acyl andalkyl being optionally substituted with one or more substituentsselected from the group consisting of acyl, alkenyl, alkoxy, alkyl,alkylamino, alkylthio, alkynyl, amino, aryl, aryloxy, aroyl, carbamate,cyano, cycloalkyl, halogen, haloalkoxy, haloalkyl, heteroalkyl,heterocycloalkyl, heteroaryl, hydrazinyl, hydroxy, mercaptyl,perhaloalkoxy, sulfonate, alkylsulfonyl, N-sulfonamido, S-sulfonamido,and thiol: if R₁ is hydrogen, methyl, —CH₂F, or fluoro, R₂, R₃, R₄, R₅,R₇, and R₈ are each hydrogen, and R₆ is hydroxyl, then R₉ is nothydrogen, formyl, unsubstituted C₁-C₅ alkylacyl, or benzoyl; if R₁ ishydrogen, methyl, —CH₂F, or fluoro, R₂, R₃, R₄, R₇, and R₈ are eachhydrogen, R₅ is methyl, and R₆ is hydroxyl, then R₉ is not hydrogen,formyl, unsubstituted C₁-C₅ alkylacyl, trifluoroacetyl, —C(O)-adamantyl,or benzoyl; if R₁ is hydrogen, methyl, fluoro, or chloro, R₂, R₃, R₄,R₆, R₇, and R₈ are each hydrogen, and R₅ is methyl, then R₉ is nothydrogen, unsubstituted C₁-C₅ alkylacyl, or benzoyl; if said dashed lineindicates a double bond, R₁, R₂, R₃, R₄, R₇, and R₈ are each hydrogen,and R₅ and R₆ are each methyl, then R₉ is not hydrogen, acetyl, orbenzoyl; if said dashed line indicates a double bond, R₁, R₂, R₃, R₄,R₅, R₇, and R₈ are each hydrogen, and R₆ is ethyl, then R₉ is notacetyl; if said dashed line does not indicate a double bond, R₁ ishydrogen or fluoro, and R₂, R₃, R₄, R₅, R₆, R₇, and R₈ are eachhydrogen, then R₉ is not hydrogen or acetyl; if R₁, R₂, R₄, R₅, R₇, andR₈ are each hydrogen, R₃ is methyl, and R₆ is hydroxyl, then R₉ is nothydrogen, formyl, unsubstituted C₁-C₅ alkylacyl, or benzoyl; and if R₁and R₂ are each fluoro, R₃, R₄, R₇, and R₈ are each hydrogen, R₅ ismethyl, and R₆ is hydroxyl, then R₉ is not acetyl.

In further embodiments, R₅ is selected from the group consisting ofC₂-C₈ alkyl, aryl, cycloalkyl, heterocycloalkyl, and heteroaryl, saidC₂-C₈ alkyl, aryl, cycloalkyl, heterocycloalkyl, and heteroaryl beingoptionally substituted with one or more substituents selected from thegroup consisting of acyl, alkenyl, alkoxy, alkyl, alkynyl, amido, amino,aryl, aryloxy, cycloalkyl, haloalkoxy, haloalkyl, heteroalkyl,heteroaryl, hydroxy, perhaloalkoxy, and thiol.

In further embodiments, R₁ and R₃ are each hydrogen; R₂ and R₄ are eachindependently selected from the group consisting of hydrogen, methyl,and fluorine; R₅ is selected from the group consisting of hydrogen,unsubstituted lower alkyl, and phenyl; R₆ is selected from the groupconsisting of hydrogen, hydroxyl, and methyl; R₇ and R₈ are eachhydrogen; and R₉ is selected from the group consisting of hydrogen,acyl, and alkyl, said acyl and alkyl being optionally substituted withamino, hydroxyl, and carboxyl.

In further embodiments, R₂ and R₄ are each hydrogen; R₅ is selected fromthe group consisting of hydrogen, unsubstituted lower alkyl, and phenyl;R₆ is selected from the group consisting of hydrogen, hydroxyl, andmethyl; R₇ and R₈ are each hydrogen; and R₉ is selected from the groupconsisting of hydrogen, acyl, and alkyl, said acyl and alkyl beingoptionally substituted with amino, hydroxyl, and carboxyl.

In further embodiments, R₅ is selected from the group consisting ofhydrogen, methyl, and ethyl; R₆ is selected from the group consisting ofhydrogen, hydroxyl, and methyl; and R₉ is selected from the groupconsisting of hydrogen, acetyl, and C(O)CH₂CH₂CO₂H.

In further embodiments, R₅ is selected from the group consisting ofunsubstituted C₂-C₆ alkyl and phenyl; R₆ is selected from the groupconsisting of hydrogen, hydroxyl, and methyl; and R₉ is selected fromthe group consisting of hydrogen, acetyl, and —C(O)CH₂CH₂CO₂H.

In further embodiments, R₅ is ethyl.

In further embodiments, said dashed line indicates an optional doublebond; R₁, R₂, R₃, and R₄ are each independently selected from the groupconsisting of hydrogen, unsubstituted lower alkyl, lower haloalkyl, andhalogen; R₅ is selected from the group consisting of hydrogen, loweralkyl, aryl, cycloalkyl, heterocycloalkyl, and heteroaryl, said loweralkyl, aryl, cycloalkyl, heterocycloalkyl, and heteroaryl beingoptionally substituted with one or more substituents selected from thegroup consisting of acyl, alkenyl, alkoxy, alkyl, alkynyl, amido, amino,aryl, aryloxy, cycloalkyl, haloalkoxy, haloalkyl, heteroalkyl,heteroaryl, hydroxy, perhaloalkoxy, and thiol; R₆ is selected from thegroup consisting of hydrogen, hydroxyl, and lower alkyl, said loweralkyl being optionally substituted with one or more substituentsselected from the group consisting of alkenyl, alkoxy, alkyl, alkynyl,aryl, aryloxy, haloalkoxy, haloalkyl, heteroalkyl, hydroxy, and thiol;R₇ and R₈ are independently selected from the groups consisting ofhydrogen, unsubstituted C₁₋₃ alkyl, or R₇ and R₈ can be taken togetherto form oxo or C₃₋₆ saturated cycloalkyl; and R₉ is selected from thegroup consisting of hydrogen, acyl, and alkyl, said acyl and alkyl beingoptionally substituted with one or more substituents selected from thegroup consisting of acyl, alkenyl, alkoxy, alkyl, alkylamino, alkylthio,alkynyl, amido, amino, aryl, aryloxy, aroyl, carbamate, carboxyl, cyano,cycloalkyl, halogen, haloalkoxy, haloalkyl, heteroalkyl,heterocycloalkyl, heteroaryl, hydrazinyl, hydroxy, mercaptyl, nitro,oxo, perhaloalkoxy, alkylsulfonyl, N-sulfonamido, S-sulfonamido, andthiol; if R₁, R₂, R₃, R₄, R₇, and R₈ are each hydrogen, R₅ is methyl,and R₆ is hydroxyl, then R₉ is not —C(O)CH₂CH₂CO₂H; if said dashed linedoes not indicate a double bond, R₁, R₂, R₃, R₄, R₅, R₇, and R₈ are eachhydrogen, and R₆ is hydroxyl, then R₉ is not —C(O)CH₂CH₂CO₂H; if saiddashed line indicates a double bond, R₁ is methyl, R₂, R₃, R₄, R₅, R₇,and R₈ are each hydrogen, and R₆ is hydroxyl, then R₉ is not—C(O)CH₂CH₂CO₂H; if R₁, R₂, R₃, R₄, R₅, R₇, and R₈ are each hydrogen andR₆ is hydroxyl, then R₉ is not hydrogen or acetyl; if said dashed linedoes not indicate a double bond, R₁ is fluorine, R₂, R₃, R₄, R₇, and R₈are each hydrogen, R₅ is methyl, and R₆ is hydroxyl, then R₉ is nothydrogen or acetyl; and if said dashed line does not indicate a doublebond, R₁ is methyl, R₂, R₃, R₄, R₅, R₇, and R₈ are each hydrogen, and R₆is hydroxyl, then R₉ is not acetyl.

In further embodiments, R₇ and R₈ are independently selected from thegroups consisting of hydrogen, unsubstituted C₂-C₃ alkyl, or R₇ and R₈can be taken together to form C₃₋₆ saturated cycloalkyl; R₉ is selectedfrom the group consisting of hydrogen, acyl, and alkyl, said acyl andalkyl being optionally substituted with one or more substituentsselected from the group consisting of acyl, alkenyl, alkoxy, alkyl,alkylamino, alkylthio, alkynyl, amino, aryl, aryloxy, aroyl, carbamate,cyano, cycloalkyl, halogen, haloalkoxy, haloalkyl, heteroalkyl,heterocycloalkyl, heteroaryl, hydrazinyl, hydroxy, mercaptyl,perhaloalkoxy, alkyl sulfonyl, N-sulfonamido, S-sulfonamido, and thiol;if R₁ is hydrogen, methyl, —CH₂F, or fluoro, R₂, R₃, R₄, R₅, R₇, and R₈are each hydrogen, and R₆ is hydroxyl, then R₉ is not hydrogen, formyl,unsubstituted C₁-C₅ alkylacyl, or benzoyl; if R₁ is hydrogen, methyl,—CH₂F, or fluoro, R₂, R₃, R₄, R₇, and R₈ are each hydrogen, R₅ ismethyl, and R₆ is hydroxyl, then R₉ is not hydrogen, formyl,unsubstituted C₁-C₅ alkylacyl, trifluoroacetyl, —C(O)-adamantyl, orbenzoyl; if R₁ is hydrogen, methyl, fluoro, or chloro, R₂, R₃, R₄, R₆,R₇, and R₈ are each hydrogen, and R₅ is methyl, then R₉ is not hydrogen,unsubstituted C₁-C₅ alkylacyl, or benzoyl; if said dashed line indicatesa double bond, R₁, R₂, R₃, R₄, R₇, and R₈ are each hydrogen, and R₅ andR₆ are each methyl, then R₉ is not hydrogen, acetyl, or benzoyl; if saiddashed line indicates a double bond, R₁, R₂, R₃, R₄, R₅, R₇, and R₈ areeach hydrogen, and R₆ is ethyl, then R₉ is not acetyl; if said dashedline does not indicate a double bond, R₁ is hydrogen or fluoro, and R₂,R₃, R₄, R₅, R₆, R₇, and R₈ are each hydrogen, then R₉ is not hydrogen oracetyl; if R₁, R₂, R₄, R₅, R₇, and R₈ are each hydrogen, R₃ is methyl,and R₆ is hydroxyl, then R₉ is not hydrogen, formyl, unsubstituted C₁-C₅alkylacyl, or benzoyl; and if R₁ and R₂ are each fluoro, R₃, R₄, R₇, andR₈ are each hydrogen, R₅ is methyl, and R₆ is hydroxyl, then R₉ is notacetyl.

In further embodiments, R₅ is selected from the group consisting ofC₂-C₈ alkyl, aryl, cycloalkyl, heterocycloalkyl, and heteroaryl, saidC₂-C₈ alkyl, aryl, cycloalkyl, heterocycloalkyl, and heteroaryl beingoptionally substituted with one or more substituents selected from thegroup consisting of acyl, alkenyl, alkoxy, alkyl, alkynyl, amido, amino,aryl, aryloxy, cycloalkyl, haloalkoxy, haloalkyl, heteroalkyl,heteroaryl, hydroxy, perhaloalkoxy, and thiol.

In further embodiments, R₁ and R₃ are each hydrogen; R₂ and R₄ are eachindependently selected from the group consisting of hydrogen, methyl,and fluorine; R₅ is selected from the group consisting of hydrogen,unsubstituted lower alkyl, and phenyl; R₆ is selected from the groupconsisting of hydrogen, hydroxyl, and methyl; R₇ and R₈ are eachhydrogen; and R₉ is selected from the group consisting of hydrogen,acyl, and alkyl, said acyl and alkyl being optionally substituted withamino, hydroxyl, and carboxyl.

In further embodiments, R₂ and R₄ are each hydrogen; R₅ is selected fromthe group consisting of hydrogen, unsubstituted lower alkyl, and phenyl;R₆ is selected from the group consisting of hydrogen, hydroxyl, andmethyl; R₇ and R₈ are each hydrogen; and R₉ is selected from the groupconsisting of hydrogen, acyl, and alkyl, said acyl and alkyl beingoptionally substituted with amino, hydroxyl, and carboxyl.

In further embodiments, R₅ is selected from the group consisting ofhydrogen, methyl, and ethyl; R₆ is selected from the group consisting ofhydrogen, hydroxyl, and methyl; and R₉ is selected from the groupconsisting of hydrogen, acetyl, and —C(O)CH₂CH₂CO₂H.

In further embodiments, R₅ is selected from the group consisting ofunsubstituted C₂-C₆ alkyl and phenyl; R₆ is selected from the groupconsisting of hydrogen, hydroxyl, and methyl; and R₉ is selected fromthe group consisting of hydrogen, acetyl, and —C(O)CH₂CH₂CO₂H.

In further embodiments, R₅ is ethyl.

In further embodiments, said dashed line indicates an optional doublebond; R₁, R₂, R₃, and R₄ are each independently selected from the groupconsisting of hydrogen, unsubstituted lower alkyl, lower haloalkyl, andhalogen; R₅ is selected from the group consisting of hydrogen, loweralkyl, aryl, cycloalkyl, heterocycloalkyl, and heteroaryl, said loweralkyl, aryl, cycloalkyl, heterocycloalkyl, and heteroaryl beingoptionally substituted with one or more substituents selected from thegroup consisting of acyl, alkenyl, alkoxy, alkyl, alkynyl, amido, amino,aryl, aryloxy, cycloalkyl, haloalkoxy, haloalkyl, heteroalkyl,heteroaryl, hydroxy, perhaloalkoxy, and thiol; R₆ is selected from thegroup consisting of hydrogen, hydroxyl, and lower alkyl, said loweralkyl being optionally substituted with one or more substituentsselected from the group consisting of alkenyl, alkoxy, alkyl, alkynyl,aryl, aryloxy, haloalkoxy, haloalkyl, heteroalkyl, hydroxy, and thiol;R₇ and R₈ are independently selected from the groups consisting ofhydrogen, unsubstituted C₂-C₃ alkyl, or R₇ and R₈ can be taken togetherto form C₃₋₆ saturated cycloalkyl; R₉ is selected from the groupconsisting of hydrogen, acyl, and alkyl, said acyl and alkyl beingoptionally substituted with one or more substituents selected from thegroup consisting of acyl, alkenyl, alkoxy, alkyl, alkylamino, alkylthio,alkynyl, amino, aryl, aryloxy, aroyl, carbamate, cyano, cycloalkyl,halogen, haloalkoxy, haloalkyl, heteroalkyl, heterocycloalkyl,heteroaryl, hydrazinyl, hydroxy, mercaptyl, perhaloalkoxy, sulfonate,alkylsulfonyl, N-sulfonamido, S-sulfonamido, and thiol; if R₁ ishydrogen, methyl, —CH₂F, or fluoro, R₂, R₃, R₄, R₅, R₇, and R₈ are eachhydrogen, and R₆ is hydroxyl, then R₉ is not hydrogen, formyl,unsubstituted C₁-C₅ alkylacyl, or benzoyl; if R₁ is hydrogen, methyl,—CH₂F, or fluoro, R₂, R₃, R₄, R₇, and R₈ are each hydrogen, R₅ ismethyl, and R₆ is hydroxyl, then R₉ is not hydrogen, formyl,unsubstituted C₁-C₅ alkylacyl, trifluoroacetyl, —C(O)-adamantyl, orbenzoyl; if R₁ is hydrogen, methyl, fluoro, or chloro, R₂, R₃, R₄, R₆,R₇, and R₈ are each hydrogen, and R₅ is methyl, then R₉ is not hydrogen,unsubstituted C₁-C₅ alkylacyl, or benzoyl; if said dashed line indicatesa double bond, R₁, R₂, R₃, R₄, R₇, and R₈ are each hydrogen, and R₅ andR₆ are each methyl, then R₉ is not hydrogen, acetyl, or benzoyl; if saiddashed line indicates a double bond, R₁, R₂, R₃, R₄, R₅, R₇, and R₈ areeach hydrogen, and R₆ is ethyl, then R₉ is not acetyl; if said dashedline does not indicate a double bond, R₁ is hydrogen or fluoro, and R₂,R₃, R₄, R₅, R₆, R₇, and R₈ are each hydrogen, then R₉ is not hydrogen oracetyl; if R₁, R₂, R₄, R₅, R₇, and R₈ are each hydrogen, R₃ is methyl,and R₆ is hydroxyl, then R₉ is not hydrogen, formyl, unsubstituted C₁-C₅alkylacyl, or benzoyl; and if R₁ and R₂ are each fluoro, R₃, R₄, R₇, andR₈ are each hydrogen, R₅ is methyl, and R₆ is hydroxyl, then R₉ is notacetyl.

In further embodiments, R₁ and R₃ are each hydrogen; R₂ and R₄ are eachindependently selected from the group consisting of hydrogen, methyl,and fluorine; R₅ is selected from the group consisting of hydrogen,unsubstituted lower alkyl, and phenyl; R₆ is selected from the groupconsisting of hydrogen, hydroxyl, and methyl; R₇ and R₈ are eachhydrogen; and R₉ is selected from the group consisting of hydrogen,acyl, and alkyl, said acyl and alkyl being optionally substituted withamino and hydroxyl.

In further embodiments, R₂ and R₄ are each hydrogen; R₅ is selected fromthe group consisting of hydrogen, unsubstituted lower alkyl, and phenyl;R₆ is selected from the group consisting of hydrogen, hydroxyl, andmethyl; R₇ and R₈ are each hydrogen; and R₉ is selected from the groupconsisting of hydrogen, acyl, and alkyl, said acyl and alkyl beingoptionally substituted with amino and hydroxyl.

In further embodiments, R₅ is selected from the group consisting ofhydrogen, methyl, and ethyl; R₆ is selected from the group consisting ofhydrogen, hydroxyl, and methyl; and R₉ is selected from the groupconsisting of hydrogen and acetyl.

In further embodiments, said dashed line indicates an optional doublebond; R₁, R₂, R₃, and R₄ are each independently selected from the groupconsisting of hydrogen, unsubstituted lower alkyl, lower haloalkyl, andhalogen; R₅ is selected from the group consisting of C₂-C₈ alkyl, aryl,cycloalkyl, heterocycloalkyl, and heteroaryl, said C₂-C₈ alkyl, aryl,cycloalkyl, heterocycloalkyl, and heteroaryl being optionallysubstituted with one or more substituents selected from the groupconsisting of acyl, alkenyl, alkoxy, alkyl, alkynyl, amido, amino, aryl,aryloxy, cycloalkyl, haloalkoxy, haloalkyl, heteroalkyl, heteroaryl,hydroxy, perhaloalkoxy, and thiol; R₆ is selected from the groupconsisting of hydrogen, hydroxyl, and lower alkyl, said lower alkylbeing optionally substituted with one or more substituents selected fromthe group consisting of alkenyl, alkoxy, alkyl, alkynyl, aryl, aryloxy,haloalkoxy, haloalkyl, heteroalkyl, hydroxy, and thiol; R₇ and R₈ areindependently selected from the groups consisting of hydrogen,unsubstituted C₁₋₃ alkyl, or R₇ and R₈ can be taken together to form oxoor C₃₋₆ saturated cycloalkyl; and R₉ is selected from the groupconsisting of hydrogen, acyl, and alkyl, said acyl and alkyl beingoptionally substituted with one or more substituents selected from thegroup consisting of acyl, alkenyl, alkoxy, alkyl, alkylamino, alkylthio,alkynyl, amido, amino, aryl, aryloxy, aroyl, carbamate, carboxyl, cyano,cycloalkyl, halogen, haloalkoxy, haloalkyl, heteroalkyl,heterocycloalkyl, heteroaryl, hydrazinyl, hydroxy, mercaptyl, nitro,oxo, perhaloalkoxy, sulfonate, alkyl sulfonyl, N-sulfonamido,S-sulfonamido, and thiol.

In further embodiments, R₁ and R₃ are each hydrogen; R₂ and R₄ are eachindependently selected from the group consisting of hydrogen, methyl,and fluorine; R₅ is selected from the group consisting of unsubstitutedC₂-C₆ alkyl and phenyl; R₆ is selected from the group consisting ofhydrogen, hydroxyl, and methyl; R₇ and R₈ are each hydrogen; and R₉ isselected from the group consisting of hydrogen, acyl, and alkyl, saidacyl and alkyl being optionally substituted with amino, hydroxyl, andcarboxyl.

In further embodiments, R₂ and R₄ are each hydrogen; R₆ is selected fromthe group consisting of hydrogen, hydroxyl, and methyl; R₇ and R₈ areeach hydrogen; and R₉ is selected from the group consisting of hydrogen,acyl, and alkyl, said acyl and alkyl being optionally substituted withamino, hydroxyl, and carboxyl.

In further embodiments, R₆ is selected from the group consisting ofhydrogen, hydroxyl, and methyl; and R₉ is selected from the groupconsisting of hydrogen, acetyl, and —C(O)CH₂CH₂CO₂H.

In further embodiments, R₅ is ethyl.

Certain compounds disclosed herein may possess useful NF-κB modulatingactivity, and may be used in the treatment or prophylaxis of a diseaseor condition in which NF-κB plays an active role. Thus, in broad aspect,certain embodiments also provide pharmaceutical compositions comprisingone or more compounds disclosed herein together with a pharmaceuticallyacceptable carrier, as well as methods of making and using the compoundsand compositions. Certain embodiments provide methods for modulatingNF-κB. Other embodiments provide methods for treating a NF-κB-mediateddisorder in a patient in need of such treatment, comprisingadministering to said patient a therapeutically effective amount of acompound or composition according to the present invention. Alsoprovided is the use of certain compounds disclosed herein for use in themanufacture of a medicament for the treatment of a disease or conditionameliorated by the modulation of NF-κB.

In certain embodiments, said NF-βB-mediated disease is selected from thegroup consisting of muscular dystrophy, arthritis, traumatic braininjury, spinal cord injury, sepsis, rheumatic disease, canceratherosclerosis, type 1 diabetes, type 2 diabetes, leptospiriosis renaldisease, glaucoma, retinal disease, ageing, headache, pain, complexregional pain syndrome, cardiac hypertrophy, muscle wasting, catabolicdisorders, obesity, fetal growth retardation, hypercholesterolemia,heart disease, chronic heart failure, ischemia/reperfusion, stroke,cerebral aneurysm, angina pectoris, pulmonary disease, cystic fibrosis,acid-induced lung injury, pulmonary hypertension, asthma, chronicobstructive pulmonary disease, Sjogren's syndrome, hyaline membranedisease, kidney disease, glomerular disease, alcoholic liver disease,gut diseases, peritoneal endometriosis, skin diseases, nasal sinusitis,mesothelioma, anhidrotic ecodermal dysplasia-ID, behcet's disease,incontinentia pigmenti, tuberculosis, asthma, crohn's disease, colitis,ocular allergy, appendicitis, paget's disease, pancreatitis,periodonitis, endometriosis, inflammatory bowel disease, inflammatorylung disease, silica-induced diseases, sleep apnea, AIDS, HIV-1,autoimmune diseases, antiphospholipid syndrome, lupus, lupus nephritis,familial mediterranean fever, hereditary periodic fever syndrome,psychosocial stress diseases, neuropathological diseases, familialamyloidotic polyneuropathy, inflammatory neuropathy, parkinson'sdisease, multiple sclerosis, alzheimer's disease, amyotropic lateralsclerosis, huntington's disease, cataracts, and hearing loss.

In further embodiments, said disease is said NF-κB-mediated disease isasthma or chronic obstructive pulmonary disease

In further embodiments, said NF-κB-mediated is Sjogren's syndrome.

In further embodiments, said NF-κB-mediated is arthritis.

In further embodiments, said NF-κB-mediated is muscular wasting.

In further embodiments, said muscular wasting disease is musculardystrophy.

In further embodiments, said muscular dystrophy is selected from thegroup consisting of Duchenne muscular dystrophy, Becker musculardystrophy, limb girdle muscular dystrophy, congenital musculardystrophy, facioscapulohumeral muscular dystrophy, myotonic musculardystrophy, oculopharyngeal muscular dystrophy, distal musculardystrophy, and Emery-Dreifuss muscular dystrophy.

In further embodiments, said muscular dystrophy is Duchenne musculardystrophy.

When ranges of values are disclosed, and the notation “from n₁ . . . ton₂” is used, where n₁ and n₂ are the numbers, then unless otherwisespecified, this notation is intended to include the numbers themselvesand the range between them. This range may be integral or continuousbetween and including the end values. By way of example, the range “from2 to 6 carbons” is intended to include two, three, four, five, and sixcarbons, since carbons come in integer units. Compare, by way ofexample, the range “from 1 to 3 μM (micromolar),” which is intended toinclude 1 μM, 3 μM, and everything in between to any number ofsignificant figures (e.g., 1.255 μM, 2.1 μM, 2.9999 μM, etc.).

The term “about,” as used herein, is intended to qualify the numericalvalues which it modifies, denoting such a value as variable within amargin of error. When no particular margin of error, such as a standarddeviation to a mean value given in a chart or table of data, is recited,the term “about” should be understood to mean that range which wouldencompass the recited value and the range which would be included byrounding up or down to that figure as well, taking into accountsignificant figures.

The term “acyl,” as used herein, alone or in combination, refers to acarbonyl attached to an alkenyl, alkyl, aryl, cycloalkyl, heteroaryl,heterocycle, or any other moiety were the atom attached to the carbonylis carbon. An “acetyl” group refers to a —C(O)CH₃ group. An“alkylcarbonyl” or “alkanoyl” group refers to an alkyl group attached tothe parent molecular moiety through a carbonyl group. Examples of suchgroups include methylcarbonyl and ethylcarbonyl. Examples of acyl groupsinclude formyl, alkanoyl and aroyl.

The term “alkenyl,” as used herein, alone or in combination, refers to astraight-chain or branched-chain hydrocarbon group having one or moredouble bonds and containing from 2 to 20 carbon atoms. In certainembodiments, said alkenyl will comprise from 2 to 6 carbon atoms. Theterm “alkenylene” refers to a carbon-carbon double bond system attachedat two or more positions such as ethenylene [(CH═CH—), (—C::C—)].Examples of suitable alkenyl groups include ethenyl, propenyl,2-methylpropenyl, 1,4-butadienyl and the like. Unless otherwisespecified, the term “alkenyl” may include “alkenylene” groups.

The term “alkoxy,” as used herein, alone or in combination, refers to analkyl ether group, wherein the term alkyl is as defined below. Examplesof suitable alkyl ether groups include methoxy, ethoxy, n-propoxy,isopropoxy, n-butoxy, iso-butoxy, sec-butoxy, tert-butoxy, and the like.

The term “alkyl,” as used herein, alone or in combination, refers to astraight-chain or branched-chain alkyl group containing from 1 to 20carbon atoms. In certain embodiments, said alkyl will comprise from 1 to10 carbon atoms. In further embodiments, said alkyl will comprise from 1to 6 carbon atoms. Alkyl groups may be optionally substituted as definedherein. Examples of alkyl groups include methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl,hexyl, octyl, noyl and the like. The term “alkylene,” as used herein,alone or in combination, refers to a saturated aliphatic group derivedfrom a straight or branched chain saturated hydrocarbon attached at twoor more positions, such as methylene (—CH₂—). Unless otherwisespecified, the term “alkyl” may include “alkylene” groups.

The term “alkylamino,” as used herein, alone or in combination, refersto an alkyl group attached to the parent molecular moiety through anamino group. Suitable alkylamino groups may be mono- or dialkylated,forming groups such as, for example, N-methylamino, N-ethylamino,N,N-dimethylamino, N,N-ethylmethylamino and the like.

The term “alkylidene,” as used herein, alone or in combination, refersto an alkenyl group in which one carbon atom of the carbon-carbon doublebond belongs to the moiety to which the alkenyl group is attached.

The term “alkylthio,” as used herein, alone or in combination, refers toan alkyl thioether (R—S—) group wherein the term alkyl is as definedabove and wherein the sulfur may be singly or doubly oxidized. Examplesof suitable alkyl thioether groups include methylthio, ethylthio,n-propylthio, isopropylthio, n-butylthio, iso-butylthio, sec-butylthio,tert-butylthio, methanesulfonyl, ethanesulfinyl, and the like.

The term “alkynyl,” as used herein, alone or in combination, refers to astraight-chain or branched chain hydrocarbon group having one or moretriple bonds and containing from 2 to 20 carbon atoms. In certainembodiments, said alkynyl comprises from 2 to 6 carbon atoms. In furtherembodiments, said alkynyl comprises from 2 to 4 carbon atoms. The term“alkynylene” refers to a carbon-carbon triple bond attached at twopositions such as ethynylene (—C:::C—, —C≡C—). Examples of alkynylgroups include ethynyl, propynyl, hydroxypropynyl, butyn-1-yl,butyn-2-yl, pentyn-1-yl, 3-methylbutyn-1-yl, hexyn-2-yl, and the like.Unless otherwise specified, the term “alkynyl” may include “alkynylene”groups.

The terms “amido” and “carbamoyl,” as used herein, alone or incombination, refer to an amino group as described below attached to theparent molecular moiety through a carbonyl group, or vice versa. Theterm “C-amido” as used herein, alone or in combination, refers to a—C(═O)—NR₂ group with R as defined herein. The term “N-amido” as usedherein, alone or in combination, refers to a RC(═O)NH— group, with R asdefined herein. The term “acylamino” as used herein, alone or incombination, embraces an acyl group attached to the parent moietythrough an amino group. An example of an “acylamino” group isacetylamino (CH₃C(O)NH—).

The term “amino,” as used herein, alone or in combination, refers to—NRR′, wherein R and R′ are independently selected from the groupconsisting of hydrogen, alkyl, acyl, heteroalkyl, aryl, cycloalkyl,heteroaryl, and heterocycloalkyl, any of which may themselves beoptionally substituted. Additionally, R and R′ may combine to formheterocycloalkyl, either of which may be optionally substituted.

The term “aryl,” as used herein, alone or in combination, means acarbocyclic aromatic system containing one, two or three rings whereinsuch polycyclic ring systems are fused together. The term “aryl”embraces aromatic groups such as phenyl, naphthyl, anthracenyl, andphenanthryl.

The term “arylalkenyl” or “aralkenyl,” as used herein, alone or incombination, refers to an aryl group attached to the parent molecularmoiety through an alkenyl group.

The term “arylalkoxy” or “aralkoxy,” as used herein, alone or incombination, refers to an aryl group attached to the parent molecularmoiety through an alkoxy group.

The term “arylalkyl” or “aralkyl,” as used herein, alone or incombination, refers to an aryl group attached to the parent molecularmoiety through an alkyl group.

The term “arylalkynyl” or “aralkynyl,” as used herein, alone or incombination, refers to an aryl group attached to the parent molecularmoiety through an alkynyl group.

The term “arylalkanoyl” or “aralkanoyl” or “aroyl,” as used herein,alone or in combination, refers to an acyl group derived from anaryl-substituted alkanecarboxylic acid such as benzoyl, napthoyl,phenylacetyl, 3-phenylpropionyl (hydrocinnamoyl), 4-phenylbutyryl,(2-naphthyl)acetyl, 4-chlorohydrocinnamoyl, and the like.

The term aryloxy as used herein, alone or in combination, refers to anaryl group attached to the parent molecular moiety through an oxy.

The terms “benzo” and “benz,” as used herein, alone or in combination,refer to the divalent group C₆H₄═ derived from benzene. Examples includebenzothiophene and benzimidazole.

The term “carbamate,” as used herein, alone or in combination, refers toan ester of carbamic acid (—NHCOO—) which may be attached to the parentmolecular moiety from either the nitrogen or acid end, and which may beoptionally substituted as defined herein.

The term “O-carbamyl” as used herein, alone or in combination, refers toa —OC(O)NRR′, group—with R and R′ as defined herein.

The term “N-carbamyl” as used herein, alone or in combination, refers toa ROC(O)NR′— group, with R and R′ as defined herein.

The term “carbonyl,” as used herein, when alone includes formyl [—C(O)H]and in combination is a —C(O)— group.

The term “carboxyl” or “carboxy,” as used herein, refers to —C(O)OH orthe corresponding “carboxylate” anion, such as is in a carboxylic acidsalt. An “O-carboxy” group refers to a RC(O)O— group, where R is asdefined herein. A “C-carboxy” group refers to a —C(O)OR groups where Ris as defined herein.

The term “cyano,” as used herein, alone or in combination, refers to—CN.

The term “cycloalkyl,” or, alternatively, “carbocycle,” as used herein,alone or in combination, refers to a saturated or partially saturatedmonocyclic, bicyclic or tricyclic alkyl group wherein each cyclic moietycontains from 3 to 12 carbon atom ring members and which may optionallybe a benzo fused ring system which is optionally substituted as definedherein. In certain embodiments, said cycloalkyl will comprise from 3 to7 carbon atoms. In certain embodiments, said cycloalkyl will comprisefrom 5 to 7 carbon atoms. Examples of such cycloalkyl groups includecyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,tetrahydronapthyl, indanyl, octahydronaphthyl, 2,3-dihydro-1H-indenyl,adamantyl and the like. “Bicyclic” and “tricyclic” as used herein areintended to include both fused ring systems, such asdecahydronaphthalene, octahydronaphthalene as well as the multicyclic(multicentered) saturated or partially unsaturated type. The latter typeof isomer is exemplified in general by, bicyclo[1,1,1]pentane, camphor,adamantane, and bicyclo[3,2,1]octane.

The term “ester,” as used herein, alone or in combination, refers to acarboxy group bridging two moieties linked at carbon atoms.

The term “ether,” as used herein, alone or in combination, refers to anoxy group bridging two moieties linked at carbon atoms.

The term “halo,” or “halogen,” as used herein, alone or in combination,refers to fluorine, chlorine, bromine, or iodine.

The term “haloalkoxy,” as used herein, alone or in combination, refersto a haloalkyl group attached to the parent molecular moiety through anoxygen atom.

The term “haloalkyl,” as used herein, alone or in combination, refers toan alkyl group having the meaning as defined above wherein one or morehydrogens are replaced with a halogen. Specifically embraced aremonohaloalkyl, dihaloalkyl and polyhaloalkyl groups. A monohaloalkylgroup, for one example, may have an iodo, bromo, chloro or fluoro atomwithin the group. Dihalo and polyhaloalkyl groups may have two or moreof the same halo atoms or a combination of different halo groups.Examples of haloalkyl groups include fluoromethyl, difluoromethyl,trifluoromethyl, chloromethyl, dichloromethyl, trichloromethyl,pentafluoroethyl, heptafluoropropyl, difluorochloromethyl,dichlorofluoromethyl, difluoroethyl, difluoropropyl, dichloroethyl anddichloropropyl. “Haloalkylene” refers to a haloalkyl group attached attwo or more positions. Examples include fluoromethylene (—CFH—),difluoromethylene (—CF₂—), chloromethylene (—CHCl—) and the like.

The term “heteroalkyl,” as used herein, alone or in combination, refersto a stable straight or branched chain, or cyclic hydrocarbon group, orcombinations thereof, fully saturated or containing from 1 to 3 degreesof unsaturation, consisting of the stated number of carbon atoms andfrom one to three heteroatoms selected from the group consisting of O,N, and S, and wherein the nitrogen and sulfur atoms may optionally beoxidized and the nitrogen heteroatom may optionally be quaternized. Theheteroatom(s) O, N and S may be placed at any interior position of theheteroalkyl group. Up to two heteroatoms may be consecutive, such as,for example, —CH₂—NH—OCH₃.

The term “heteroaryl,” as used herein, alone or in combination, refersto a 3 to 7 membered unsaturated heteromonocyclic ring, or a fusedmonocyclic, bicyclic, or tricyclic ring system in which at least one ofthe fused rings is aromatic, which contains at least one atom selectedfrom the group consisting of O, S, and N. In certain embodiments, saidheteroaryl will comprise from 5 to 7 carbon atoms. The term alsoembraces fused polycyclic groups wherein heterocyclic rings are fusedwith aryl rings, wherein heteroaryl rings are fused with otherheteroaryl rings, wherein heteroaryl rings are fused withheterocycloalkyl rings, or wherein heteroaryl rings are fused withcycloalkyl rings. Examples of heteroaryl groups include pyrrolyl,pyrrolinyl, imidazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl,pyridazinyl, triazolyl, pyranyl, furyl, thienyl, oxazolyl, isoxazolyl,oxadiazolyl, thiazolyl, thiadiazolyl, isothiazolyl, indolyl, isoindolyl,indolizinyl, benzimidazolyl, quinolyl, isoquinolyl, quinoxalinyl,quinazolinyl, indazolyl, benzotriazolyl, benzodioxolyl, benzopyranyl,benzoxazolyl, benzoxadiazolyl, benzothiazolyl, benzothiadiazolyl,benzofuryl, benzothienyl, chromonyl, coumarinyl, benzopyranyl,tetrahydroquinolinyl, tetrazolopyridazinyl, tetrahydroisoquinolinyl,thienopyridinyl, furopyridinyl, pyrrolopyridinyl and the like. Exemplarytricyclic heterocyclic groups include carbazolyl, benzindolyl,phenanthrolinyl, dibenzofuranyl, acridinyl, phenanthridinyl, xanthenyland the like.

The terms “heterocycloalkyl” and, interchangeably, “heterocycle,” asused herein, alone or in combination, each refer to a saturated,partially unsaturated, or fully unsaturated monocyclic, bicyclic, ortricyclic heterocyclic group containing at least one heteroatom as aring member, wherein each said heteroatom may be independently selectedfrom the group consisting of nitrogen, oxygen, and sulfur In certainembodiments, said heterocycloalkyl will comprise from 1 to 4 heteroatomsas ring members. In further embodiments, said heterocycloalkyl willcomprise from 1 to 2 heteroatoms as ring members. In certainembodiments, said heterocycloalkyl will comprise from 3 to 8 ringmembers in each ring. In further embodiments, said heterocycloalkyl willcomprise from 3 to 7 ring members in each ring. In yet furtherembodiments, said heterocycloalkyl will comprise from 5 to 6 ringmembers in each ring. “Heterocycloalkyl” and “heterocycle” are intendedto include sulfones, sulfoxides, N-oxides of tertiary nitrogen ringmembers, and carbocyclic fused and benzo fused ring systems;additionally, both terms also include systems where a heterocycle ringis fused to an aryl group, as defined herein, or an additionalheterocycle group. Examples of heterocycle groups include aziridinyl,azetidinyl, 1,3-benzodioxolyl, dihydroisoindolyl, dihydroisoquinolinyl,dihydrocinnolinyl, dihydrobenzodioxinyl,dihydro[1,3]oxazolo[4,5-b]pyridinyl, benzothiazolyl, dihydroindolyl,dihy-dropyridinyl, 1,3-dioxanyl, 1,4-dioxanyl, 1,3-dioxolanyl,isoindolinyl, morpholinyl, piperazinyl, pyrrolidinyl,tetrahydropyridinyl, piperidinyl, thiomorpholinyl, and the like. Theheterocycle groups may be optionally substituted unless specificallyprohibited.

The term “hydrazinyl” as used herein, alone or in combination, refers totwo amino groups joined by a single bond, i.e., —N—N—.

The term “hydroxy,” as used herein, alone or in combination, refers to—OH.

The term “hydroxyalkyl,” as used herein, alone or in combination, refersto a hydroxy group attached to the parent molecular moiety through analkyl group.

The term “imino,” as used herein, alone or in combination, refers to═N—.

The term “iminohydroxy,” as used herein, alone or in combination, refersto ═N(OH) and ═N—O—.

The phrase “in the main chain” refers to the longest contiguous oradjacent chain of carbon atoms starting at the point of attachment of agroup to the compounds of any one of the formulas disclosed herein.

The term “isocyanato” refers to a —NCO group.

The term “isothiocyanato” refers to a —NCS group.

The phrase “linear chain of atoms” refers to the longest straight chainof atoms independently selected from carbon, nitrogen, oxygen andsulfur.

The term “lower,” as used herein, alone or in a combination, where nototherwise specifically defined, means containing from 1 to and including6 carbon atoms.

The term “lower aryl,” as used herein, alone or in combination, meansphenyl or naphthyl, which may be optionally substituted as provided.

The term “lower heteroaryl,” as used herein, alone or in combination,means either 1) monocyclic heteroaryl comprising five or six ringmembers, of which between one and four said members may be heteroatomsselected from the group consisting of O, S, and N, or 2) bicyclicheteroaryl, wherein each of the fused rings comprises five or six ringmembers, comprising between them one to four heteroatoms selected fromthe group consisting of O, S, and N.

The term “lower cycloalkyl,” as used herein, alone or in combination,means a monocyclic cycloalkyl having between three and six ring members.Lower cycloalkyls may be unsaturated. Examples of lower cycloalkylinclude cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

The term “lower heterocycloalkyl,” as used herein, alone or incombination, means a monocyclic heterocycloalkyl having between threeand six ring members, of which between one and four may be heteroatomsselected from the group consisting of O, S, and N. Examples of lowerheterocycloalkyls include pyrrolidinyl, imidazolidinyl, pyrazolidinyl,piperidinyl, piperazinyl, and morpholinyl. Lower heterocycloalkyls maybe unsaturated.

The term “lower amino,” as used herein, alone or in combination, refersto —NRR′, wherein R and R′ are independently selected from the groupconsisting of hydrogen, lower alkyl, and lower heteroalkyl, any of whichmay be optionally substituted. Additionally, the R and R′ of a loweramino group may combine to form a five- or six-memberedheterocycloalkyl, either of which may be optionally substituted.

The term “mercaptyl” as used herein, alone or in combination, refers toan RS— group, where R is as defined herein.

The term “nitro,” as used herein, alone or in combination, refers to—NO₂.

The terms “oxy” or “oxa,” as used herein, alone or in combination, referto —O—.

The term “oxo,” as used herein, alone or in combination, refers to ═O.

The term “perhaloalkoxy” refers to an alkoxy group where all of thehydrogen atoms are replaced by halogen atoms.

The term “perhaloalkyl” as used herein, alone or in combination, refersto an alkyl group where all of the hydrogen atoms are replaced byhalogen atoms.

The terms “sulfonate,” “sulfonic acid,” and “sulfonic,” as used herein,alone or in combination, refer the SO₃H group and its anion as thesulfonic acid is used in salt formation.

The term “sulfanyl,” as used herein, alone or in combination, refers to—S—.

The term “sulfinyl,” as used herein, alone or in combination, refers to—S(O)—.

The term “sulfonyl,” as used herein, alone or in combination, refers to—S(O)₂—.

The term “N-sulfonamido” refers to a RS(═O)₂NR′— group with R and R′ asdefined herein.

The term “S-sulfonamido” refers to a —S(═O)₂NRR′, group, with R and R′as defined herein.

The terms “thia” and “thio,” as used herein, alone or in combination,refer to a —S— group or an ether wherein the oxygen is replaced withsulfur. The oxidized derivatives of the thio group, namely sulfinyl andsulfonyl, are included in the definition of thia and thio.

The term “thiol,” as used herein, alone or in combination, refers to an—SH group.

The term “thiocarbonyl,” as used herein, when alone includes thioformyl—C(S)H and in combination is a —C(S)— group.

The term “N-thiocarbamyl” refers to an ROC(S)NR′ group, with R and R′ asdefined herein.

The term “O-thiocarbamyl” refers to a —OC(S)NRR′, group with R and R′ asdefined herein.

The term “thiocyanato” refers to a —CNS group.

The term “trihalomethanesulfonamido” refers to a X₃CS(O)₂NR— group withX is a halogen and R as defined herein.

The term “trihalomethanesulfonyl” refers to a X₃CS(O)₂— group where X isa halogen.

The term “trihalomethoxy” refers to a X₃CO— group where X is a halogen.

The term “trisubstituted silyl,” as used herein, alone or incombination, refers to a silicone group substituted at its three freevalences with groups as listed herein under the definition ofsubstituted amino. Examples include trimethysilyl,tert-butyldimethylsilyl, triphenylsilyl and the like.

Any definition herein may be used in combination with any otherdefinition to describe a composite structural group. By convention, thetrailing element of any such definition is that which attaches to theparent moiety. For example, the composite group alkylamido wouldrepresent an alkyl group attached to the parent molecule through anamido group, and the term alkoxyalkyl would represent an alkoxy groupattached to the parent molecule through an alkyl group.

When a group is defined to be “null,” what is meant is that said groupis absent.

The term “optionally substituted” means the anteceding group may besubstituted or unsubstituted. When substituted, the substituents of an“optionally substituted” group may include, without limitation, one ormore substituents independently selected from the following groups or aparticular designated set of groups, alone or in combination: loweralkyl, lower alkenyl, lower alkynyl, lower alkanoyl, lower heteroalkyl,lower heterocycloalkyl, lower haloalkyl, lower haloalkenyl, lowerhaloalkynyl, lower perhaloalkyl, lower perhaloalkoxy, lower cycloalkyl,phenyl, aryl, aryloxy, lower alkoxy, lower haloalkoxy, oxo, loweracyloxy, carbonyl, carboxyl, lower alkylcarbonyl, lower carboxyester,lower carboxamido, cyano, hydrogen, halogen, hydroxy, amino, loweralkylamino, arylamino, amido, nitro, thiol, lower alkylthio, lowerhaloalkylthio, lower perhaloalkylthio, arylthio, sulfonate, sulfonicacid, trisubstituted silyl, N₃, SH, SCH₃, C(O)CH₃, CO₂CH₃, CO₂H,pyridinyl, thiophene, furanyl, lower carbamate, and lower urea. Twosubstituents may be joined together to form a fused five-, six-, orseven-membered carbocyclic or heterocyclic ring consisting of zero tothree heteroatoms, for example forming methylenedioxy or ethylenedioxy.An optionally substituted group may be unsubstituted (e.g., —CH₂CH₃),fully substituted (e.g., —CF₂CF₃), monosubstituted (e.g., —CH₂CH₂F) orsubstituted at a level anywhere in-between fully substituted andmonosubstituted (e.g., —CH₂CF₃). Where substituents are recited withoutqualification as to substitution, both substituted and unsubstitutedforms are encompassed. Where a substituent is qualified as“substituted,” the substituted form is specifically intended.Additionally, different sets of optional substituents to a particularmoiety may be defined as needed; in these cases, the optionalsubstitution will be as defined, often immediately following the phrase,“optionally substituted with.”

The term R or the term R′, appearing by itself and without a numberdesignation, unless otherwise defined, refers to a moiety selected fromthe group consisting of hydrogen, alkyl, cycloalkyl, heteroalkyl, aryl,heteroaryl and heterocycloalkyl, any of which may be optionallysubstituted. Such R and R′ groups should be understood to be optionallysubstituted as defined herein. Whether an R group has a numberdesignation or not, every R group, including R, R′ and R^(n) where n=(1,2, 3, . . . n), every substituent, and every term should be understoodto be independent of every other in terms of selection from a group.Should any variable, substituent, or term (e.g. aryl, heterocycle, R,etc.) occur more than one time in a formula or generic structure, itsdefinition at each occurrence is independent of the definition at everyother occurrence. Those of skill in the art will further recognize thatcertain groups may be attached to a parent molecule or may occupy aposition in a chain of elements from either end as written. Thus, by wayof example only, an unsymmetrical group such as —C(O)N(R)— may beattached to the parent moiety at either the carbon or the nitrogen.

Asymmetric centers exist in the compounds disclosed herein. Thesecenters are designated by the symbols “R” or “S,” depending on theconfiguration of substituents around the chiral carbon atom. It shouldbe understood that the invention encompasses all stereochemical isomericforms, including diastereomeric, enantiomeric, and epimeric forms, aswell as d-isomers and 1-isomers, and mixtures thereof. Individualstereoisomers of compounds can be prepared synthetically fromcommercially available starting materials which contain chiral centersor by preparation of mixtures of enantiomeric products followed byseparation such as conversion to a mixture of diastereomers followed byseparation or recrystallization, chromatographic techniques, directseparation of enantiomers on chiral chromatographic columns, or anyother appropriate method known in the art. Starting compounds ofparticular stereochemistry are either commercially available or can bemade and resolved by techniques known in the art. Additionally, thecompounds disclosed herein may exist as geometric isomers. The presentinvention includes all cis, trans, syn, anti, entgegen (E), and zusammen(Z) isomers as well as the appropriate mixtures thereof. Additionally,compounds may exist as tautomers; all tautomeric isomers are provided bythis invention. Additionally, the compounds disclosed herein can existin unsolvated as well as solvated forms with pharmaceutically acceptablesolvents such as water, ethanol, and the like. In general, the solvatedforms are considered equivalent to the unsolvated forms.

The term “bond” refers to a covalent linkage between two atoms, or twomoieties when the atoms joined by the bond are considered to be part oflarger substructure. A bond may be single, double, or triple unlessotherwise specified. A dashed line between two atoms in a drawing of amolecule indicates that an additional bond may be present or absent atthat position.

The term “disease” as used herein is intended to be generallysynonymous, and is used interchangeably with, the terms “disorder” and“condition” (as in medical condition), in that all reflect an abnormalcondition of the human or animal body or of one of its parts thatimpairs normal functioning, is typically manifested by distinguishingsigns and symptoms, and causes the human or animal to have a reducedduration or quality of life.

The term “NF-κB-mediated disease,” refers to a disease in which NF-κBplays an active role in the disease pathology. NF-κB-mediated diseasesinclude diseases in which multiple biological pathways and/or processesin addition to NF-κB-mediated processes contribute to the diseasepathology. A NF-κB-mediated disease may be completely or partiallymediated by modulating the activity or amount of NF-κB. In particular, aNF-κB-mediated disease is one in which modulation of NF-κB results insome effect on the underlying disease e.g., administration of a NF-κBmodulator results in some improvement in at least some of the patientsbeing treated. The term “NF-κB-mediated disease” also refers to thefollowing diseases, even though the compounds disclosed herein exerttheir effects through biological pathways and/or processes other thanNF-κB: muscular dystrophy, arthritis, traumatic brain injury, spinalcord injury, sepsis, rheumatic disease, cancer atherosclerosis, type 1diabetes, type 2 diabetes, leptospiriosis renal disease, glaucoma,retinal disease, ageing, headache, pain, complex regional pain syndrome,cardiac hypertrophy, muscle wasting, catabolic disorders, obesity, fetalgrowth retardation, hypercholesterolemia, heart disease, chronic heartfailure, ischemia/reperfusion, stroke, cerebral aneurysm, anginapectoris, pulmonary disease, cystic fibrosis, acid-induced lung injury,pulmonary hypertension, asthma, chronic obstructive pulmonary disease,Sjogren's syndrome, hyaline membrane disease, kidney disease, glomerulardisease, alcoholic liver disease, gut diseases, peritonealendometriosis, skin diseases, nasal sinusitis, mesothelioma, anhidroticecodermal dysplasia-ID, behcet's disease, incontinentia pigmenti,tuberculosis, asthma, crohn's disease, colitis, ocular allergy,appendicitis, paget's disease, pancreatitis, periodonitis,endometriosis, inflammatory bowel disease, inflammatory lung disease,silica-induced diseases, sleep apnea, AIDS, HIV-1, autoimmune diseases,antiphospholipid syndrome, lupus, lupus nephritis, familialmediterranean fever, hereditary periodic fever syndrome, psychosocialstress diseases, neuropathological diseases, familial amyloidoticpolyneuropathy, inflammatory neuropathy, parkinson's disease, multiplesclerosis, alzheimer's disease, amyotropic lateral sclerosis,huntington's disease, cataracts, and hearing loss.

The term “combination therapy” means the administration of two or moretherapeutic agents to treat a therapeutic condition or disorderdescribed in the present disclosure. Such administration encompassesco-administration of these therapeutic agents in a substantiallysimultaneous manner, such as in a single capsule having a fixed ratio ofactive ingredients or in multiple, separate capsules for each activeingredient. In addition, such administration also encompasses use ofeach type of therapeutic agent in a sequential manner. In either case,the treatment regimen will provide beneficial effects of the drugcombination in treating the conditions or disorders described herein.

“NF-κB modulator is used herein to refer to a compound that exhibits anEC₅₀ with respect to NF-κB activity of no more than about 100 μM andmore typically not more than about 50 μM, as measured in the NF-κBinhibitor assays described generally hereinbelow. “EC₅₀” is thatconcentration of modulator which either activates or reduces theactivity or increases or decreases the amount of an enzyme (e.g.,(NF-κB)) to half-maximal level. Certain compounds disclosed herein havebeen discovered to exhibit modulatory activity against NF-κB. In certainembodiments, compounds will exhibit an EC₅₀ with respect to NF-κB of nomore than about 10 μM; in further embodiments, compounds will exhibit anEC₅₀ with respect to NF-κB of no more than about 5 μM; in yet furtherembodiments, compounds will exhibit an EC₅₀ with respect to NF-κB of notmore than about 1 μM; in yet further embodiments, compounds will exhibitan EC₅₀ with respect to NF-κB of not more than about 200 nM, as measuredin the NF-κB assay described herein.

The phrase “therapeutically effective” is intended to qualify the amountof active ingredients used in the treatment of a disease or disorder.This amount will achieve the goal of reducing or eliminating the saiddisease or disorder.

The term “therapeutically acceptable” refers to those compounds (orsalts, prodrugs, tautomers, zwitterionic forms, etc.) which are suitablefor use in contact with the tissues of patients without undue toxicity,irritation, and allergic response, are commensurate with a reasonablebenefit/risk ratio, and are effective for their intended use.

As used herein, reference to “treatment” of a patient is intended toinclude prophylaxis. The term “patient” means all mammals includinghumans. Examples of patients include humans, cows, dogs, cats, goats,sheep, pigs, and rabbits. Preferably, the patient is a human.

The term “prodrug” refers to a compound that is made more active invivo. Certain compounds disclosed herein may also exist as prodrugs, asdescribed in Hydrolysis in Drug and Prodrug Metabolism: Chemistry,Biochemistry, and Enzymology (Testa, Bernard and Mayer, Joachim M.Wiley-VHCA, Zurich, Switzerland 2003). Prodrugs of the compoundsdescribed herein are structurally modified forms of the compound thatreadily undergo chemical changes under physiological conditions toprovide the compound. Additionally, prodrugs can be converted to thecompound by chemical or biochemical methods in an ex vivo environment.For example, prodrugs can be slowly converted to a compound when placedin a transdermal patch reservoir with a suitable enzyme or chemicalreagent. Prodrugs are often useful because, in some situations, they maybe easier to administer than the compound, or parent drug. They may, forinstance, be bioavailable by oral administration whereas the parent drugis not. The prodrug may also have improved solubility in pharmaceuticalcompositions over the parent drug. A wide variety of prodrug derivativesare known in the art, such as those that rely on hydrolytic cleavage oroxidative activation of the prodrug. An example, without limitation, ofa prodrug would be a compound which is administered as an ester (the“prodrug”), but then is metabolically hydrolyzed to the carboxylic acid,the active entity. Additional examples include peptidyl derivatives of acompound.

The compounds disclosed herein can exist as therapeutically acceptablesalts. The present invention includes compounds listed above in the formof salts, including acid addition salts. Suitable salts include thoseformed with both organic and inorganic acids. Such acid addition saltswill normally be pharmaceutically acceptable. However, salts ofnon-pharmaceutically acceptable salts may be of utility in thepreparation and purification of the compound in question. Basic additionsalts may also be formed and be pharmaceutically acceptable. For a morecomplete discussion of the preparation and selection of salts, refer toPharmaceutical Salts: Properties, Selection, and Use (Stahl, P.Heinrich. Wiley-VCHA, Zurich, Switzerland, 2002).

The terms “therapeutically acceptable salt,” or “salt,” as used herein,represents salts or zwitterionic forms of the compounds disclosed hereinwhich are water or oil-soluble or dispersible and therapeuticallyacceptable as defined herein. The salts can be prepared during the finalisolation and purification of the compounds or separately by reactingthe appropriate compound in the form of the free base with a suitableacid. Representative acid addition salts include acetate, adipate,alginate, L-ascorbate, aspartate, benzoate, benzenesulfonate (besylate),bisulfate, butyrate, camphorate, camphorsulfonate, citrate, digluconate,formate, fumarate, gentisate, glutarate, glycerophosphate, glycolate,hemisulfate, heptanoate, hexanoate, hippurate, hydrochloride,hydrobromide, hydroiodide, 2-hydroxyethansulfonate (isethionate),lactate, maleate, malonate, DL-mandelate, mesitylenesulfonate,methanesulfonate, naphthylenesulfonate, nicotinate,2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate,3-phenylproprionate, phosphonate, picrate, pivalate, propionate,pyroglutamate, succinate, sulfonate, tartrate, L-tartrate,trichloroacetate, trifluoroacetate, phosphate, glutamate, bicarbonate,para-toluenesulfonate (p-tosylate), and undecanoate. Also, basic groupsin the compounds disclosed herein can be quaternized with methyl, ethyl,propyl, and butyl chlorides, bromides, and iodides; dimethyl, diethyl,dibutyl, and diamyl sulfates; decyl, lauryl, myristyl, and sterylchlorides, bromides, and iodides; and benzyl and phenethyl bromides.Examples of acids which can be employed to form therapeuticallyacceptable addition salts include inorganic acids such as hydrochloric,hydrobromic, sulfuric, and phosphoric, and organic acids such as oxalic,maleic, succinic, and citric. Salts can also be formed by coordinationof the compounds with an alkali metal or alkaline earth ion. Hence, thepresent invention contemplates sodium, potassium, magnesium, and calciumsalts of the compounds disclosed herein, and the like.

Basic addition salts can be prepared during the final isolation andpurification of the compounds by reacting a carboxy group with asuitable base such as the hydroxide, carbonate, or bicarbonate of ametal cation or with ammonia or an organic primary, secondary, ortertiary amine. The cations of therapeutically acceptable salts includelithium, sodium, potassium, calcium, magnesium, and aluminum, as well asnontoxic quaternary amine cations such as ammonium, tetramethylammonium,tetraethylammonium, methylamine, dimethylamine, trimethylamine,triethylamine, diethylamine, ethylamine, tributylamine, pyridine,N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine,dicyclohexylamine, procaine, dibenzylamine, N,N-dibenzylphenethylamine,1-ephenamine, and N,N-dibenzylethylenediamine. Other representativeorganic amines useful for the formation of base addition salts includeethylenediamine, ethanolamine, diethanolamine, piperidine, andpiperazine.

In certain embodiments, the salts may include hydrochloride,hydrobromide, sulfonate, citrate, tartrate, phosphonate, lactate,pyruvate, acetate, succinate, oxalate, fumarate, malate, oxaloacetate,methanesulfonate, ethanesulfonate, p-toluenesulfonate, benzenesulfonateand isethionate salts of compounds disclosed herein. A salt of acompound can be made by reacting the appropriate compound in the form ofthe free base with the appropriate acid.

While it may be possible for the compounds of the subject invention tobe administered as the raw chemical, it is also possible to present themas a pharmaceutical formulation. Accordingly, provided herein arepharmaceutical formulations which comprise one or more of certaincompounds disclosed herein, or one or more pharmaceutically acceptablesalts, esters, prodrugs, amides, or solvates thereof, together with oneor more pharmaceutically acceptable carriers thereof and optionally oneor more other therapeutic ingredients. The carrier(s) must be“acceptable” in the sense of being compatible with the other ingredientsof the formulation and not deleterious to the recipient thereof. Properformulation is dependent upon the route of administration chosen. Any ofthe well-known techniques, carriers, and excipients may be used assuitable and as understood in the art; e.g., in Remington'sPharmaceutical Sciences. The pharmaceutical compositions disclosedherein may be manufactured in any manner known in the art, e.g., bymeans of conventional mixing, dissolving, granulating, dragee-making,levigating, emulsifying, encapsulating, entrapping or compressionprocesses.

The formulations include those suitable for oral, parenteral (includingsubcutaneous, intradermal, intramuscular, intravenous, intraarticular,and intramedullary), intraperitoneal, transmucosal, transdermal, rectaland topical (including dermal, buccal, sublingual and intraocular)administration although the most suitable route may depend upon forexample the condition and disorder of the recipient. The formulationsmay conveniently be presented in unit dosage form and may be prepared byany of the methods well known in the art of pharmacy. Typically, thesemethods include the step of bringing into association a compound of thesubject invention or a pharmaceutically acceptable salt, ester, amide,prodrug or solvate thereof (“active ingredient”) with the carrier whichconstitutes one or more accessory ingredients. In general, theformulations are prepared by uniformly and intimately bringing intoassociation the active ingredient with liquid carriers or finely dividedsolid carriers or both and then, if necessary, shaping the product intothe desired formulation.

Formulations of the compounds disclosed herein suitable for oraladministration may be presented as discrete units such as capsules,cachets or tablets each containing a predetermined amount of the activeingredient; as a powder or granules; as a solution or a suspension in anaqueous liquid or a non-aqueous liquid; or as an oil-in-water liquidemulsion or a water-in-oil liquid emulsion. The active ingredient mayalso be presented as a bolus, electuary or paste.

Pharmaceutical preparations which can be used orally include tablets,push-fit capsules made of gelatin, as well as soft, sealed capsules madeof gelatin and a plasticizer, such as glycerol or sorbitol. Tablets maybe made by compression or molding, optionally with one or more accessoryingredients. Compressed tablets may be prepared by compressing in asuitable machine the active ingredient in a free-flowing form such as apowder or granules, optionally mixed with binders, inert diluents, orlubricating, surface active or dispersing agents. Molded tablets may bemade by molding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent. The tablets may optionally becoated or scored and may be formulated so as to provide slow orcontrolled release of the active ingredient therein. All formulationsfor oral administration should be in dosages suitable for suchadministration. The push-fit capsules can contain the active ingredientsin admixture with filler such as lactose, binders such as starches,and/or lubricants such as talc or magnesium stearate and, optionally,stabilizers. In soft capsules, the active compounds may be dissolved orsuspended in suitable liquids, such as fatty oils, liquid paraffin, orliquid polyethylene glycols. In addition, stabilizers may be added.Dragee cores are provided with suitable coatings. For this purpose,concentrated sugar solutions may be used, which may optionally containgum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethyleneglycol, and/or titanium dioxide, lacquer solutions, and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments may be added to thetablets or dragee coatings for identification or to characterizedifferent combinations of active compound doses.

The compounds may be formulated for parenteral administration byinjection, e.g., by bolus injection or continuous infusion. Formulationsfor injection may be presented in unit dosage form, e.g., in ampoules orin multi-dose containers, with an added preservative. The compositionsmay take such forms as suspensions, solutions or emulsions in oily oraqueous vehicles, and may contain formulatory agents such as suspending,stabilizing and/or dispersing agents. The formulations may be presentedin unit-dose or multi-dose containers, for example sealed ampoules andvials, and may be stored in powder form or in a freeze-dried(lyophilized) condition requiring only the addition of the sterileliquid carrier, for example, saline or sterile pyrogen-free water,immediately prior to use. Extemporaneous injection solutions andsuspensions may be prepared from sterile powders, granules and tabletsof the kind previously described.

Formulations for parenteral administration include aqueous andnon-aqueous (oily) sterile injection solutions of the active compoundswhich may contain antioxidants, buffers, bacteriostats and solutes whichrender the formulation isotonic with the blood of the intendedrecipient; and aqueous and non-aqueous sterile suspensions which mayinclude suspending agents and thickening agents. Suitable lipophilicsolvents or vehicles include fatty oils such as sesame oil, or syntheticfatty acid esters, such as ethyl oleate or triglycerides, or liposomes.Aqueous injection suspensions may contain substances which increase theviscosity of the suspension, such as sodium carboxymethyl cellulose,sorbitol, or dextran. Optionally, the suspension may also containsuitable stabilizers or agents which increase the solubility of thecompounds to allow for the preparation of highly concentrated solutions.

For oral or parenteral use, the compounds may be formulated asnanoparticle preparations. Such nanoparticle preparations can include,for example, nanoshere encapsulations of active compounds, inactivenanoparticles to which active compounds can be tethered, or nanoscalepowders of active compounds. Nanoparticle preparations can be used toincrease the bioavailability of the active compounds, control the rateof release of the active compounds, or deliver active compounds to aparticular location in the body. See A. Dove, “An Easy Pill to Swallow”,Drug Discovery & Development Magazine: 11(11), November 2008, pp. 22-24.

In addition to the formulations described previously, the compounds mayalso be formulated as a depot preparation. Such long acting formulationsmay be administered by implantation (for example subcutaneously orintramuscularly) or by intramuscular injection. Thus, for example, thecompounds may be formulated with suitable polymeric or hydrophobicmaterials (for example as an emulsion in an acceptable oil) or ionexchange resins, or as sparingly soluble derivatives, for example, as asparingly soluble salt.

For buccal or sublingual administration, the compositions may take theform of tablets, lozenges, pastilles, or gels formulated in conventionalmanner. Such compositions may comprise the active ingredient in aflavored basis such as sucrose and acacia or tragacanth.

The compounds may also be formulated in rectal compositions such assuppositories or retention enemas, e.g., containing conventionalsuppository bases such as cocoa butter, polyethylene glycol, or otherglycerides.

Certain compounds disclosed herein may be administered topically, thatis by non-systemic administration. This includes the application of acompound disclosed herein externally to the epidermis or the buccalcavity and the instillation of such a compound into the ear, eye andnose, such that the compound does not significantly enter the bloodstream. In contrast, systemic administration refers to oral,intravenous, intraperitoneal and intramuscular administration.

Formulations suitable for topical administration include liquid orsemi-liquid preparations suitable for penetration through the skin tothe site of inflammation such as gels, liniments, lotions, creams,ointments or pastes, and drops suitable for administration to the eye,ear or nose. The active ingredient for topical administration maycomprise, for example, from 0.001% to 10% w/w (by weight) of theformulation. In certain embodiments, the active ingredient may compriseas much as 10% w/w. In other embodiments, it may comprise less than 5%w/w. In certain embodiments, the active ingredient may comprise from 2%w/w to 5% w/w. In other embodiments, it may comprise from 0.1% to 1% w/wof the formulation.

Formulations for topical administration in the mouth, for examplebuccally or sublingually, include lozenges comprising the activeingredient in a flavored basis such as sucrose and acacia or tragacanth,and pastilles comprising the active ingredient in a basis such asgelatin and glycerin or sucrose and acacia.

For administration by inhalation, compounds may be convenientlydelivered from an insufflator, nebulizer pressurized packs or otherconvenient means of delivering an aerosol spray. Pressurized packs maycomprise a suitable propellant such as dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas. In the case of a pressurized aerosol, the dosageunit may be determined by providing a valve to deliver a metered amount.Alternatively, for administration by inhalation or insufflation, thecompounds according to the invention may take the form of a dry powdercomposition, for example a powder mix of the compound and a suitablepowder base such as lactose or starch. The powder composition may bepresented in unit dosage form, in for example, capsules, cartridges,gelatin or blister packs from which the powder may be administered withthe aid of an inhalator or insufflator.

Preferred unit dosage formulations are those containing an effectivedose, as herein below recited, or an appropriate fraction thereof, ofthe active ingredient.

It should be understood that in addition to the ingredients particularlymentioned above, the formulations described above may include otheragents conventional in the art having regard to the type of formulationin question, for example those suitable for oral administration mayinclude flavoring agents.

Compounds may be administered orally or via injection at a dose of from0.1 to 500 mg/kg per day. The dose range for adult humans is generallyfrom 5 mg to 2 g/day. Tablets or other forms of presentation provided indiscrete units may conveniently contain an amount of one or morecompounds which is effective at such dosage or as a multiple of thesame, for instance, units containing 5 mg to 500 mg, usually around 10mg to 200 mg.

The amount of active ingredient that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration.

The compounds can be administered in various modes, e.g. orally,topically, or by injection. The precise amount of compound administeredto a patient will be the responsibility of the attendant physician. Thespecific dose level for any particular patient will depend upon avariety of factors including the activity of the specific compoundemployed, the age, body weight, general health, sex, diets, time ofadministration, route of administration, rate of excretion, drugcombination, the precise disorder being treated, and the severity of theindication or condition being treated. Also, the route of administrationmay vary depending on the condition and its severity.

In certain instances, it may be appropriate to administer at least oneof the compounds described herein (or a pharmaceutically acceptablesalt, ester, or prodrug thereof) in combination with another therapeuticagent. By way of example only, if one of the side effects experienced bya patient upon receiving one of the compounds herein is hypertension,then it may be appropriate to administer an anti-hypertensive agent incombination with the initial therapeutic agent. Or, by way of exampleonly, the therapeutic effectiveness of one of the compounds describedherein may be enhanced by administration of an adjuvant (i.e., by itselfthe adjuvant may only have minimal therapeutic benefit, but incombination with another therapeutic agent, the overall therapeuticbenefit to the patient is enhanced). Or, by way of example only, thebenefit of experienced by a patient may be increased by administeringone of the compounds described herein with another therapeutic agent(which also includes a therapeutic regimen) that also has therapeuticbenefit. By way of example only, in a treatment for diabetes involvingadministration of one of the compounds described herein, increasedtherapeutic benefit may result by also providing the patient withanother therapeutic agent for diabetes. In any case, regardless of thedisease, disorder or condition being treated, the overall benefitexperienced by the patient may simply be additive of the two therapeuticagents or the patient may experience a synergistic benefit.

In any case, the multiple therapeutic agents (at least one of which is acompound disclosed herein) may be administered in any order or evensimultaneously. If simultaneously, the multiple therapeutic agents maybe provided in a single, unified form, or in multiple forms (by way ofexample only, either as a single pill or as two separate pills). One ofthe therapeutic agents may be given in multiple doses, or both may begiven as multiple doses. If not simultaneous, the timing between themultiple doses may be any duration of time ranging from a few minutes tofour weeks.

Thus, in another aspect, certain embodiments provide methods fortreating NF-κB-mediated disorders in a human or animal subject in needof such treatment comprising administering to said subject an amount ofa compound disclosed herein effective to reduce or prevent said disorderin the subject, in combination with at least one additional agent forthe treatment of said disorder that is known in the art. In a relatedaspect, certain embodiments provide therapeutic compositions comprisingat least one compound disclosed herein in combination with one or moreadditional agents for the treatment of NF-κB-mediated disorders.

Specific diseases to be treated by the compounds, compositions, andmethods disclosed herein include ageing, headache, pain, complexregional pain syndrome, cardiac hypertrophy, muscular dystrophy, musclewasting, catabolic disorders, type 1 diabetes, type 2 diabetes, obesity,fetal growth retardation, hypercholesterolemia, atherosclerosis, heartdisease, chronic heart failure, ischemia/reperfusion, stroke, cerebralaneurysm, angina pectoris, pulmonary disease, cystic fibrosis,acid-induced lung injury, pulmonary hypertension, chronic obstructivepulmonary disease, hyaline membrane disease, kidney disease, glomerulardisease, alcoholic liver disease, leptospiriosis renal disease, gutdiseases, peritoneal endometriosis, skin diseases, nasal sinusitis,mesothelioma, anhidrotic ecodermal dysplasia-ID, behcet's disease,incontinentia pigmenti, tuberculosis, asthma, arthritis, crohn'sdisease, colitis, ocular allergy, glaucoma, appendicitis, paget'sdisease, pancreatitis, periodonitis, endometriosis, inflammatory boweldisease, inflammatory lung disease, sepsis, silica-induced diseases,sleep apnea, AIDS, HIV-1, autoimmune diseases, antiphospholipidsyndrome, lupus, lupus nephritis, familial mediterranean fever,hereditary periodic fever syndrome, psychosocial stress diseases,neuropathological diseases, familial amyloidotic polyneuropathy,inflammatory neuropathy, traumatic brain injury, spinal cord injury,parkinson's disease, multiple sclerosis, rheumatic disease, alzheimer'sdisease, amyotropic lateral sclerosis, huntington's disease, retinaldisease, cataracts, hearing loss, and cancer.

Besides being useful for human treatment, certain compounds andformulations disclosed herein may also be useful for veterinarytreatment of companion animals, exotic animals and farm animals,including mammals, rodents, and the like. More preferred animals includehorses, dogs, and cats.

All references, patents or applications, U.S. or foreign, cited in theapplication are hereby incorporated by reference as if written herein intheir entireties. Where any inconsistencies arise, material literallydisclosed herein controls.

General Synthetic Methods for Preparing Compounds

The following schemes can be used to practice the present invention.

Examples 6-7, 23, and 28 can be synthesized according to scheme I.

Examples 8 and 24-25 can be synthesized according to scheme II.

Examples 15, 22, and 27 can be synthesized according to scheme III.

Examples 19, 21, and 26 can be synthesized according to scheme IV.

Example 32 can be synthesized according to scheme V.

Examples 1-3 can be synthesized according to scheme VI.

Preparation 1 can be synthesized according to scheme VII.

Example 16 can be synthesized according to scheme VIII.

The invention is further illustrated by the following examples. AllIUPAC names were generated using CambridgeSoft's ChemDraw 10.0.

Preparation 12-oxo-2-((6S,10R,13S)-6,10,13-trimethyl-3-oxo-6,7,8,10,12,13,14,15-octahydro-3H-cyclopenta[a]phenanthren-17-yl)ethylacetate

Step 1

2-((6S,10R,13S)-17-hydroxy-6,10,13-trimethyl-3-oxo-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-17-yl)-2-oxoethylacetate: (see Tetrahedron Letters, 2001, 42 (14): 2639-2642).Alternatively, methyl prednisolone 21-acetate is dissolved in a mixtureof dimethylformamide and tetrahydrofuran and cooled in an ice bath. SO₂is bubbled into methanesulfonyl chloride and the mixture is addeddropwise to the solution containing the solution of methyl prednisolone21-acetate. The title product can then be isolated by standard aqueousworkup.

Step 2

(2′R,4′R,6S,10R,13S)-2′-acetyl-2′,6,10,13-tetramethyl-7,8,10,12,13,14,15,16-octahydrospiro[cyclopenta[a]phenanthrene-17,4′-[1,3]dioxane]-3,5′(6H)-dione:2-((6S,10R,13S)-17-hydroxy-6,10,13-trimethyl-3-oxo-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-17-yl)-2-oxoethylacetate is dissolved in toluene and heated with 1.5 equivalents of ethylorthoacetate and a trace of pyridinium hydrochloride. Ethanol isdistilled off the reaction mixture to drive it to completion.

Step 3

(6S,10R,13S)-17-(2-hydroxyacetyl)-6,10,13-trimethyl-3-oxo-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-17-ylacetate: The reaction mixture from step 3 is concentrated, dissolved intetrahydrofuran, and treated with dilute hydrochloric acid. Standardaqueous workup yields the title compound.

Step 4

2-oxo-2-((6S,10R,13S)-6,10,13-trimethyl-3-oxo-6,7,8,10,12,13,14,15-octahydro-3H-cyclopenta[a]phenanthren-17-yl)ethylacetate:(6S,10R,13S)-17-(2-hydroxyacetyl)-6,10,13-trimethyl-3-oxo-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-17-ylacetate is heated with 2 equivalents of potassium carbonate in dimethylformamide. Standard aqueous workup yields the title compound.

EXAMPLE 1(10S,13S,17R)-17-hydroxy-17-(2-hydroxyacetyl)-10,13-dimethyl-6,7,8,10,12,13,14,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-3(2H)-one

Step 1

(10S,13S,17R)-17-hydroxy-17-(2-hydroxyacetyl)-10,13-dimethyl-6,7,8,10,12,13,14,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-3(2H)-one:Commercially available as Anecortave acetate. The title compound can besynthesized according to the procedure of Example 8, Step 2,substituting2-((10S,13S,17R)-17-hydroxy-10,13-dimethyl-3-oxo-2,3,6,7,8,10,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl)-2-oxoethylacetate for2-oxo-2-((10S,13S,16R,17S)-10,13,16-trimethyl-3-oxo-2,3,6,7,8,10,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl)ethylacetate.

EXAMPLE 2(10S,13S,17R)-17-hydroxy-17-(2-hydroxyacetyl)-10,13-dimethyl-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-3-one

(10S,13S,17R)-17-hydroxy-17-(2-hydroxyacetyl)-10,13-dimethyl-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-3-one:The title compound can be synthesized according to the procedures ofExample 3, Step 1 and Example 1, Step 1, substituting prednisoloneacetate for hydrocortisone acetate.

EXAMPLE 32-((10S,13S,17R)-17-hydroxy-10,13-dimethyl-3-oxo-2,3,6,7,8,10,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl)-2-oxoethylacetate

Step 1

2-((10S,13S,17R)-17-hydroxy-10,13-dimethyl-3-oxo-2,3,6,7,8,10,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl)-2-oxoethylacetate: The title compound can be synthesized from hydrocortisoneacetate according to the procedure disclosed in EP 0097328.405 g (1 mol)of hydrocortisone acetate is added to a mixture of 2 liters ofN,N-dimethylformamide and 350 ml of pyridine, and with stirring at roomtemperature, 260 g of methanesulfonyl chloride is added. The reactionmixture is heated, maintained at 80 to 85° C. for 1 hour, and thencooled to room temperature. Methanol (7 liters) is added. Theprecipitated crystals are separated by filtration, washed with methanoland water, and dried under reduced pressure to give the title compound.

EXAMPLE 6(10S,13S,16R,17S)-17-(2-hydroxyacetyl)-10,13,16-trimethyl-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-3-one

Step 1

(10S,13S,16R,17S)-17-(2-hydroxyacetyl)-10,13,16-trimethyl-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-3-one:A solution of the product from Example 7, step 2 in methylene chlorideand methanol (1:3 methylene chloride/methanol) is stirred under an inertatmosphere and cooled in an ice bath. Aqueous potassium carbonate isadded by syringe. The reaction is stirred at 5° C. for 2 hours. Thereaction is then neutralized with 1N HCl and concentrated. Afterpartitioning between water and methylene chloride, the product solutionis dried over anhydrous magnesium sulfate, filtered and evaporated togive the title compound.

EXAMPLE 72-oxo-2-((10S,13S,16R)-10,13,16-trimethyl-3-oxo-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-17-yl)ethylacetate

Step 1

2-((10S,13S,16R)-10,13,16-trimethyl-3-oxo-7,8,12,13,15,16-hexahydro-3H-cyclopenta[a]phenanthren-17(6H,10H,14H)-ylidene)-2-(trimethylsilyloxy)ethylacetate: (see K. P. Shephard, U.S. Pat. No. 4,975,536; Dec. 4, 1990;Preparation 1, col. 8) Into predried reactor 1 was added 36.64 grams(100 mmole) of2-((10S,13S)-10,13-dimethyl-3-oxo-6,7,8,10,12,13,14,15-octahydro-3H-cyclopenta[a]phenanthren-17-yl)-2-oxoethylacetate (commercial product from Pfizer). The starting material wasdissolved in 200 ml of anhydrous tetrahydrofuran and 200 ml of anhydrousdichloromethane. Trimethylsilyl imidazole, (20.0 ml, 136 mmole), wasadded. This solution was cooled to −50° C. under a small nitrogen flow.

Into predried reactor 2 was added copper II propionate (2.10 grams, 10.0mmole), 150 ml of anhydrous tetrahydrofuran, and anhydrous1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone. The mixture wascooled to −50° C. and methyl magnesium chloride (3M, 10.0 ml) was addeddropwise over approximately 5 minutes. The mixture was stirred forapproximately 10 minutes. The contents of reactor 2 were transferred toreactor 1 via cannula quickly (approximately 30 sec.), and reactor 2 wasrinsed with 10 ml of anhydrous tetrahydrofuran and this was alsocannulated into reactor 1. A pump was set up with methyl magnesiumchloride (3M, 45.0 ml) and pumped into reactor 1 over 45 min (pumpsetting at 1.0 ml/min). Reactor 1 was stirred further at −50° C. for 1hour, then warmed to −30° C. overnight.

Toluene (1 L) was added and the temperature brought to 0° C. The mixturewas extracted with 2×500 ml of 5% acetic acid (cold), then with 200 mlof 25% sodium chloride. The aqueous phases were back extracted with 300ml of toluene. The combined toluene extracts were dried over magnesiumsulfate, filtered, and concentrated to a viscous oil. Yield—57.8 grams.

Step 2

2-oxo-2-((10S,13S,16R,17S)-10,13,16-trimethyl-3-oxo-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-17-yl)ethylacetate: The crude product from step 1 is dissolved in ethyl acetate,and slurried with aqueous 1N HCl until hydrolysis is complete. Theaqueous acid is neutralized with aqueous potassium bicarbonate, and theethyl acetate phase is dried, filtered, and concentrated to asemi-solid.

EXAMPLE 8(10S,13S,16R,17S)-17-(2-hydroxyacetyl)-10,13,16-trimethyl-6,7,8,10,12,13,14,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-3(2H)-one

Step 1

2-oxo-2-((10S,13S,16R,17S)-10,13,16-trimethyl-3-oxo-2,3,6,7,8,10,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl)ethylacetate: A mixture of 3.3 g (8.6 mM) of2-oxo-2-((10S,13S,16R,17S)-10,13,16-trimethyl-3-oxo-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-17-yl)ethylacetate, chlorotris(triphenylphosphine)rhodium(I) (Wilkinson's Catalyst,480 mg, 0.52 mM), triethylsilane (1.4 mL, 1.0 g, 8.8 mM) and methylenechloride (15 mL) was warmed to 40° C. and stirred until most of thestarting material was gone, as determined by thin layer chromatography.The reaction was evaporated in vacuo and chromatographed on fine silicagel (600 g) in 10-15% ethyl acetate in methylene chloride. One fractionof 700 mL was collected, followed by twelve 200 mL fractions. A 1.0 gquantity of desired product (30% yield) was obtained by evaporation offractions 6-12. (Starting material was obtained from fraction 13, 0.7 g,20% recovery). NMR (500 MHz, CDCl₃, TMS): δ 0.68 (s, 3H), 0.98 (d, 3H,J=6.5 Hz), 1.12 (m, 1H), 1.33 (s, 3H), 1.47 (m, 1H), 1.57 (m, 1H), 1.69(m, 1H), 1.99 (m, 1H), 2.18 (s, 3H), 2.07-2.29 (m, 6H), 2.36 (d, 1H),2.50 (m, 3H), 2.79 (m, 1H), 4.48 (d, 1H, J=17 Hz), 4.73 (d, 1H, J=17Hz), 5.50 (s, 1H), 5.75 (s, 1H).

Step 2

(10S,13S,16R,17S)-17-(2-hydroxyacetyl)-10,13,16-trimethyl-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-3-one:A solution of2-oxo-2-((10S,13S,16R,17S)-10,13,16-trimethyl-3-oxo-2,3,6,7,8,10,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl)ethylacetate (1.0 g, 2.6 mM) in methylene chloride (5 mL) and methanol (15mL) was put under an inert atmosphere and cooled in an ice bath. 1 mL of1 M aqueous potassium carbonate was added by syringe. The reaction wasstirred at 5° C. for 2 h. The reaction was then neutralized with 1N HCland concentrated. After partitioning between water and methylenechloride, the product solution was dried over anhydrous magnesiumsulfate, filtered and evaporated. Crystallization from ethyl acetateyielded a first crop of 0.33 g product. NMR (500 MHz, CDCl₃, TMS): δ0.67 (s, 3H), 1.01 (d, 3H, J=7 Hz), 1.13 (m, 1H), 1.33 (s, 3H),1.47-1.80 (m, 3H), 2.00 (m, 1H), 2.06-2.24 (m, 6H), 2.37 (d, 1H),2.45-2.60 (m, 3H), 2.82 (m, 1H), 3.30 (m, 1H), 4.20 (m, 2H), 5.50 (d,1H, J=5 Hz), 5.76 (s, 1H).

EXAMPLE 15(10S,13S,16R,17R)-17-hydroxy-17-(2-hydroxyacetyl)-10,13,16-trimethyl-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-3-one

Step 1

2-((10S,13S,16R,17R)-17-hydroxy-10,13,16-trimethyl-3-oxo-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-17-yl)-2-oxoethylacetate:(Z)-2-((10S,13S,16R)-10,13,16-trimethyl-3-oxo-7,8,12,13,15,16-hexahydro-3H-cyclopenta[a]phenanthren-17(6H,10H,14H)-ylidene)-2-(trimethylsilyloxy)ethylacetate is dissolved in methylene chloride and the mixture is cooled tozero degrees Celsius. A solution of m-chloroperbenzoic acid in methylenechloride is added dropwise and the mixture is stirred for 4 hours. Theorganic phase was washed with aqueous acetic acid and then aqueousbisulfite. The organic phase was concentrated and chromatographed onsilica gel to yield the title compound.

Step 2

(10S,13S,16R,17R)-17-hydroxy-17-(2-hydroxyacetyl)-10,13,16-trimethyl-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-3-one:Prepared according to Example 8, Step 2 substituting2-((10S,13S,16R,17R)-17-hydroxy-10,13,16-trimethyl-3-oxo-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-17-yl)-2-oxoethylacetate for2-oxo-2-((10S,13S,16R,17S)-10,13,16-trimethyl-3-oxo-2,3,6,7,8,10,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl)ethylacetate.

EXAMPLE 16(10S,13S,16S,17R)-17-hydroxy-17-(2-hydroxyacetyl)-10,13,16-trimethyl-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-3-one

EXAMPLE 19(10S,13S,16R,17R)-17-hydroxy-17-(2-hydroxyacetyl)-10,13,16-trimethyl-6,7,8,10,12,13,14,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-3(2H)-one

EXAMPLE 20(10S,13S,16S,17R)-17-hydroxy-17-(2-hydroxyacetyl)-10,13,16-trimethyl-6,7,8,10,12,13,14,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-3(2H)-one

EXAMPLE 21(10S,13S,16R,17R)-17-hydroxy-17-(2-hydroxyacetyl)-10,13-dimethyl-16-propyl-6,7,8,10,12,13,14,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-3(2H)-one

EXAMPLE 22(10S,13S,16R,17R)-17-hydroxy-17-(2-hydroxyacetyl)-10,13-dimethyl-16-propyl-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-3-one

EXAMPLE 23(10S,13S,16R,17S)-17-(2-hydroxyacetyl)-10,13-dimethyl-16-propyl-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-3-one

EXAMPLE 24(10S,13S,16R,17S)-17-(2-hydroxyacetyl)-10,13-dimethyl-16-propyl-6,7,8,10,12,13,14,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-3(2H)-one

EXAMPLE 25(10S,13S,16R,17S)-17-(2-hydroxyacetyl)-10,13-dimethyl-16-phenyl-6,7,8,10,12,13,14,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-3(2H)-one

EXAMPLE 26(10S,13S,16S,17R)-17-hydroxy-17-(2-hydroxyacetyl)-10,13-dimethyl-16-phenyl-6,7,8,10,12,13,14,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-3(2H)-one

EXAMPLE 27(10S,13S,16S,17R)-17-hydroxy-17-(2-hydroxyacetyl)-10,13-dimethyl-16-phenyl-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-3-one

EXAMPLE 28(10S,13S,16R,17S)-17-(2-hydroxyacetyl)-10,13-dimethyl-16-phenyl-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-3-one

EXAMPLE 29(10S,13S,16S,17S)-17-(2-hydroxyacetyl)-10,13,16-trimethyl-6,7,8,10,12,13,14,15,16,17-decahydro-3H-cyclopenta[a]phenanthren-3-one

EXAMPLE 30(10S,13S,16S,17S)-17-(2-hydroxyacetyl)-10,13,16-trimethyl-6,7,8,10,12,13,14,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-3(2H)-one

EXAMPLE 312-oxo-2-((10S,13S,16S,17S)-10,13,16-trimethyl-3-oxo-2,3,6,7,8,10,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl)ethylacetate

EXAMPLE 32(10S,13S,16R,17S)-17-(2-hydroxyacetyl)-10,13,16,17-tetramethyl-6,7,8,10,12,13,14,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-3(2H)-one

Step 1

2-oxo-2-((10S,13S,16R,17S)-10,13,16,17-tetramethyl-3-oxo-2,3,6,7,8,10,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl)ethylacetate: A mixture of2-((10S,13S)-10,13-dimethyl-3-oxo-2,3,6,7,8,10,12,13,14,15-decahydro-1H-cyclopenta[a]phenanthren-17-yl)-2-oxoethylacetate (150 g) and copper propionate (1.9M in THF (90 ml) is cooled inan ice acetone bath. Methyl magnesium chloride (1.96M in THF, 240 ml) isadded dropwise for 30 min. After 1 hour, the reaction is quenched withmethyl iodide (100 g) in 200 ml THF. The reaction mixture is thenpartitioned with water and toluene. The separated organic phase iswashed with water, dried over sodium sulfate and concentrated. Theresidue is crystallized from ether and hexane to give the titlecompound.

Step 2

10S,13S,16R,17S)-17-(2-hydroxyacetyl)-10,13,16,17-tetramethyl-6,7,8,10,12,13,14,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-3(2H)-one:2-oxo-2-((10S,13S,16R,17S)-10,13,16,17-tetramethyl-3-oxo-2,3,6,7,8,10,12,13,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-17-yl)ethylacetate (144 g) is stirred in 1500 ml methanol and treated with sodiummethoxide (25%, 5 ml) for 30 minutes. The mixture is partitioned betweenmethylene chloride and sodium bicarbonate. The organic phase isseparated and washed with sodium bicarbonate, dried over sodium sulfate,and concentrated to give the title compound.

The following compounds can generally be made using the methodsdescribed above. It is expected that these compounds when made will haveactivity similar to those that have been made and tested.

The activity of the compounds in Examples 1-7 and 15-16 as NF-κBmodulators is illustrated in the following assays. The other compoundslisted above, which have not yet been made and/or tested, are predictedto have activity in these assays as well.

Biological Activity Assay In Vitro NF-kb Inhibitor Screening Assay

C2C12 skeletal muscle cells stably transfected with a luciferasereporter construct regulated under multiple copies of the NF-kB responseelement (Panomics, Fremont, Calif.) were used to screen NF-kBinhibitors. These cells were maintained at 37° C. with 5% CO₂ in atissue culture incubator with Dulbecco's modified Eagle medium (DMEM)medium containing 10% Fetal bovine serum (FBS) (ATCC, Manassas, Va.),Penicillin 100 U/ml, Streptomycin 100 μg/ml, and 100 μg/ml Hygromysin B(Roche, Indianapolis, Ind.). Screening assays were performed inmyoblasts (grown in medium containing 10% FBS) in duplicate 96 wellplates at a cell concentration of 5×10⁴ cells per well in 100 ul volume.Cells were pretreated with various concentrations (0.01 ug/ml to 10ug/ml) of compound for 24 hr duration before stimulating with tumornecrosis factor-α (TNF-α) (10 ng/ml) for another 24 hrs. Prednisolonewas included in every plate tested as a positive control. After thecompletion of incubation cells were washed twice with PBS and lysed withcell lysis buffer to measure luciferase activity (Promega Corp, Madison,Wis.) using Centro LB 960 luminometer (Berthold technologies, GmbH & Co,Bad Wildbad, Germany). Relative luminescence units with TNF-αstimulation in the absence of drugs were considered as 100% percent anddata was represented as % inhibition relative to TNF-α induced NF-kBactivation.

Some of the compounds disclosed herein were tested in the C2C12 skeletalmuscle cell luciferase assay and exhibited ≧100% inhibition atconcentrations of 0.01, 0.1, and 1 ug/mL; 80-100% inhibition atconcentrations of 0.01, 0.1, 1, and 10 ug/mL; 60-80% inhibition atconcentrations of 0.01, 0.1, and 1 ug/mL; 40-60% inhibition atconcentrations of 1 and 10 ug/mL; and 20-40% inhibition atconcentrations of 10 ug/mL.

Cell viability was assayed in duplicate plates by MTT(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) (Sigma,Saint Louis, Mo.) as per manufacturer's protocols. Percent cellviability was calculated relative to untreated cells. There was not asignificant decrease in cell viability (<80%) for any of compoundstested at any of the doses (0.01, 0.1, 1, and 10 ug/mL) tested.

Inhibition of NF-kB Nuclear Translocation

Inhibition of TNF-α induced NF-kB activation was confirmed by nucleartranslocation immunofluorescence assay. C2C12 cells were grown on coverslips and treated with TNF-α and compound at optimal concentrations asdescribed above. Cells were fixed with acetone and stained with a rabbitanti-NF-kB (p60) antibody/anti-rabbit Texas red (Santa Cruz Biotech,Inc, Santa Cruz, and CA) and counterstained with4′,6-Diamidino-2-phenylindole HCl (DAPI) (Invitrogen, CA) to visualizethe nuclei. Some of the compounds disclosed herein were tested in thenuclear translocation immunofluorescence assay and blocked TNF-α inducedNF-kB nuclear translocation.

In Vivo mdx Mouse Model of Dystrophy

Separate groups (n=12-14) of mdx mice were treated with prednisolone (5mg/kg/day; per oral in feed), Example 1 (20 and 40 mg/kg/day; per oralin feed) and Example 2 (20 and 40 mg/kg/day; per oral in feed) for 3months. All mice underwent 30 min biweekly treadmill exercise during thetreatment duration to unmask the mild disease phenotype of mdx mousemodel.

Effect on Body Weight (BM)

Prednisolone treated mice showed significantly lower body weight(P<0.05) than untreated mice at 33.8 weeks age. Mice treated with someof the compounds disclosed herein at 20 and 40 mg/kg/day dosages gainedsignificantly more body weight and gastrocnemius muscle mass thanuntreated mice.

Effect on In Vivo Motor Coordination and Strength

Motor coordination and strength were assessed using Rota-rod (UgoBasile, VA, Italy) testing. Briefly, mice were trained on the rota-rodfor two days before collecting data. Each acclimatization sessionconsisted of four training sessions, 2 per day and each session lasting120 seconds at a speed of 5 rpm). Each trial consisted of placing themice on the rod at 10 rpm for 60 seconds (stabilizing period) followedby an acceleration from 10 rpm to 40 rpm within the first 25 secondsuntil the animal fell from the rod or until 180 seconds are reached. Ifthe animals fell during the stabilizing period, they were placed back onthe rod to complete the session. The total testing time was 240 seconds(60 seconds stabilization time and 180 seconds test time). Each trialwas performed twice a day (2 hour interval between sessions) for 3consecutive days. The latency to fall (seconds) was recorded and all sixscores were averaged per mouse. The average data was expressed aslatency to fall (in seconds) for each group mice at 3 age groups. Theability of untreated mice to stay on the rod did not changesignificantly with time. Mice treated with some of the compoundsdisclosed herein at 20 and 40 mg/kg/day dosages showed increased latencyto fall at 12, 24, or 36 weeks. In some instances, latency to fall wasdecreased or unchanged, increased by 0-10%, increased by 10-20%,increased by 20-30%, or increased by 30-40%.

Effect on In Vitro Force Contractions:

The distal tendon of the extensor digitorum longus (EDL) muscle was tiedsecurely to the lever arm of a servomotor/force transducer (model 305B,Aurora Scientific, Richmond Hill, ON, Canada) and the proximal tendon toa tissue clamp. Muscles were stimulated between two platinum electrodes.With supramaximal stimulation of the muscle using single 0.2-ms squarestimulation pulses for the EDL, muscle length was adjusted to the length(L_(o)) that resulted in maximal twitch force. With the muscle held atL_(o) using stimulation frequencies of 30, 50, 80, 100, 120 and 150 Hz,the maximum isometric tetanic force (P_(o)) developed during a 300 mstrain of stimulation pulses was recorded for the EDL muscle. The musclelength was then measured with calipers and after removal of the musclefrom the bath the mass of the muscle was determined. For each muscle,the optimum fiber length (L_(f)) was calculated by multiplying L_(o) bya previously determined L_(f)/L_(o) ratio of 0.45. Total muscle fibercross-sectional area was determined by dividing the wet mass by theproduct of L_(f) and the density of mammalian skeletal muscle (1.06mg/mm³). Maximum isometric specific force (sP_(o)) was determined bydividing P_(o) by the total muscle fiber cross-sectional area. There wasno statistically significant (P<0.05) difference in specific forcebetween untreated and prednisolone groups. Mice treated with some of thecompounds disclosed herein at 20 and 40 mg/kg/day dosages showedstatistically significant (P<0.05) increased isometric specific force.In some instances, isometric specific force was decreased or unchanged,increased by 0-5%, increased by 5-10%, increased by 10-15%, or increasedby 15-20%.

Histological Evaluations

Hematoxylin and Eosin staining of gastrocnemius muscle of untreated mdxmice show significant degeneration and inflammation. Skeletal musclefrom Example 1- and Example 2-treated mice showed significant decreasein inflammation, degeneration, and increase in regenerating musclefibers in comparison to untreated and prednisolone treated mdx mice.Continuous administration of prednisolone appeared to increasedegeneration and decrease in regeneration of dystrophic skeletal muscle.

Glucocorticoid Receptor Binding Assay

To determine the receptor binding affinity of example compounds to theglucocorticoid receptor (GR), a ligand binding assay was performed usingcDNA expression clones (Baculovirus) for human and mouse glucocorticoidreceptor-alpha. Liver extracts containing different GR constructs wereincubated with radiolabeled 3H-Dexamethsone (Amersham Pharmacia Biotech)and test compound in assay buffer (10 mM Tris-HCl, 1.5 mM EDTA, 10%glycerol, 1 mM dithiothreitol, and 20 mM sodium molybdate, pH 7.6). Theamount of radioactivity was measured using a scintillation plate reader.Dexamethasone showed competitive binding with 3H-dexamethasone atmicromolar concentrations. Some of the compounds disclosed herein weretested in the glucocorticoid receptor binding assay and found to have nosignificant (>75%) competitive binding to the glucocorticoid receptor atmillimolar concentrations.

From the foregoing description, one skilled in the art can easilyascertain the essential characteristics of this invention, and withoutdeparting from the spirit and scope thereof, can make various changesand modifications of the invention to adapt it to various usages andconditions.

1. A method of reducing the symptoms of arthritis comprising theadministration, to a patient in need thereof, of a therapeuticallyeffective amount of a compound having the structural formula:


2. A method of treating arthritis comprising the administration, to apatient in need thereof, of a therapeutically effective amount of acompound having the structural formula: